Farnesoid x receptor modulators

ABSTRACT

The present application provides a compound of formula I: 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt, solvate, or amino acid conjugate thereof, wherein R 1 -R 10 , m, n, p, and   are as described herein. The present invention relates generally to FXR modulators and to methods of making and using said compounds.

BACKGROUND

Farnesoid X receptor (FXR) is a nuclear receptor that functions as abile acid sensor controlling bile acid homeostasis. FXR is expressed invarious organs and shown to be involved in many diseases and conditions,such as liver diseases, lung diseases, renal diseases, intestinaldiseases, and heart diseases, and biological processes, includingglucose metabolism, insulin metabolism, and lipid metabolism. A numberof natural bile acids are FXR modulators, and are able to regulateFXR-mediated diseases and conditions (Gioiello, et al., 2014 Curr. Top.Med. Chem. 14, 2159). For example, natural bile acids such aschenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid(LCA), and the taurine and glycine conjugates thereof serve as FXRligands.

Derivatives of natural bile acids have also been described as FXRmodulators. European Patent No. 0312867 describes 6-methyl derivativesof natural biliary acids such as ursodeoxycholic acid, ursocholic acid,chenodeoxycholic acid and cholic acid. WO 2002/75298 discloses3α,7α-dihydroxy-6α-ethyl-5β-cholan-24-oic acid (hereinafter alsoreferred to as 6-ethyl-chenodeoxycholic acid, or 6-ECDCA), salts,solvates, and amino acid conjugates thereof as FXR modulators, which canbe used to prevent or treat FXR-mediated diseases or conditions.

However, it is well known that natural bile acids and bile acidderivatives modulate not only other nuclear hormone receptors, but arealso modulators for the G protein-coupled receptor (GPCR) TGR5. Receptorselectivity is a problem in connection with the development of atherapeutic compound directed to modulating a nuclear hormone receptorsuch as FXR. A non-selective therapeutic compound may carry an increasedrisk of side effects. Other obstacles to overcome in the development ofa therapeutic compound include a non-suitable pharmacokinetic profile,safety issues such as toxicity (e.g., liver) and undesirable drug-druginteractions.

Thus, there remains a need for additional selective FXR modulatorssuitable for drug development, for example, a compound that is selectiveagainst other nuclear receptors and/or does not significantly activatethe bile acid GPCR TGR5.

SUMMARY

An objective of the present invention is to provide compounds thatmodulate FXR. In one aspect, the present invention provides a compoundof formula I:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein:

R¹ is OH, alkoxy, halogen, or oxo;

R² and R³ are each independently H, OH, halogen, or alkyl optionallysubstituted with one or more halogen or OH, or R² and R³ taken togetherwith the carbon atom to which they are attached form a carbonyl;

R⁴ is H, halogen, alkyl optionally substituted with one or more halogenor OH, alkenyl, or alkynyl;

R⁵ and R⁶ are each independently H, OH, OSO₃H, OCOCH₃, OPO₃H₂, orhalogen, or R⁵ and R⁶ taken together with the carbon atom to which theyare attached form a carbonyl;

R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂, SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, C(O)NHOH,tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or 2,4-difluoro-3-hydroxyphenyl;

R⁸, R⁹, and R¹⁰ are each independently H, OH, halogen, or alkyloptionally substituted with one or more halogen or OH, or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2heteroatoms selected from N, O, and S, or R⁹ and R¹⁰ taken together withthe carbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S;

m is 0, 1, or 2;

n is 0 or 1;

p is 0 or 1; and

is a single or double bond, provided that when each

is a single bond, the sum of m, n, and p is 2, R¹ is OH, and R⁸, R⁹, andR¹⁰ are each H, then R⁷ is not CO₂H.

The present invention further provides a pharmaceutical compositioncomprising a compound of formula I or a pharmaceutically acceptablesalt, solvate, or amino acid conjugate thereof, and a pharmaceuticallyacceptable carrier or excipient.

The present invention also provides a method for treating or preventinga disease or condition mediated by FXR, comprising administering to asubject in need thereof an effective amount of a compound of formula Ior a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

The present invention also provides for the manufacture of a medicamentfor treating or preventing a disease or condition mediated by FXR,wherein the medicament comprises a compound of formula I or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

The present invention further provides compositions, includingpharmaceutical compositions, for use in treating or preventing a diseaseor condition mediated by FXR, wherein the composition comprises acompound of formula I or a pharmaceutically acceptable salt, solvate, oramino acid conjugate thereof.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. The materials, methods, and examples areillustrative only and are not intended to be limiting.

Other features and advantages of the invention will be apparent from thefollowing detailed description.

DETAILED DESCRIPTION Definitions

Certain terms used in the specification and claims are collected here.

As used herein, the phrase “a compound of the invention” refers to acompound of any one of formula I, II, III, IV, V, VI, VII, Ia, Ib, Ic,Id, Ie, Id, or any compound explicitly disclosed herein.

As used herein, the term “alkyl” refers to a straight-chain or branchedsaturated hydrocarbon moiety. The term “C₁-C₆ alkyl” refers to astraight-chain or branched hydrocarbon moiety having 1, 2, 3, 4, 5, or 6carbon atoms. “C₁-C₄ alkyl” refers to a straight-chain or branchedhydrocarbon moiety having 1, 2, 3, or 4 carbon atoms, including methyl,ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and t-butyl.

The term “alkenyl” refers to a straight-chain or branched hydrocarbonmoiety containing at least one carbon-carbon double bond. Both the transand cis isomers of the carbon-carbon double bond are encompassed underthe term “alkenyl”. Examples of alkenyl moieties include, but are notlimited to, vinyl, allyl, 1-butenyl, 2-butenyl, 3-butenyl, and2-hexenyl.

As used herein, “alkynyl” refers to a straight-chain or branchedhydrocarbon moiety containing at least one carbon-carbon triple bond.Examples of alkynyl moieties include, but are not limited to, ethynyl,2-propynyl, 5-but-1-en-3-ynyl, and 3-hexynyl.

The term “alkoxy” refers to a straight-chain or branched saturatedhydrocarbon covalently attached to an oxygen atom. Examples of alkoxymoieties include, but are not limited to, methoxy, ethoxy, isopropyloxy,n-propoxy, n-butoxy, t-butoxy, and pentoxy.

As used herein, the term “halogen” refers to fluorine, bromine, chlorineand iodine.

The term “optionally substituted” refers to the indicated moiety whichmay or may not be substituted, and when substituted is mono-, di-, ortri-substituted, such as with 1, 2, or 3 substituents. In someinstances, the substituent is halogen or OH.

As used herein, “carbocycle”, “carbocyclic” or “carbocyclic ring” isintended to include any stable monocyclic or bicyclic ring having thespecified number of carbons, any of which may be saturated, unsaturated,or aromatic. Carbocyclic ring includes cycloalkyl and aryl. For example,a C₃ -C₈ carbocyclic ring is intended to include a monocyclic orbicyclic ring having 3,4, 5, 6, 7, or 8 carbon atoms. Examples ofcarbocycles include, but are not limited to, cyclopropyl, cyclobutyl,cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptenyl,cycloheptyl, cycloheptenyl, adamantyl, cyclooctyl, cyclooctenyl, andphenyl.

As used herein, “heterocycle”, “heterocyclic” or “heterocyclic group”includes any ring structure (saturated, unsaturated, or aromatic) whichcontains at least one ring heteroatom (e.g., N, O or S). Heterocycleincludes heterocycloalkyl and heteroaryl. Examples of heterocyclesinclude, but are not limited to, morpholine, pyrrolidine,tetrahydrothiophene, piperidine, piperazine, oxetane, pyran,tetrahydropyran, azetidine, and tetrahydrofuran. Examples ofheterocyclic groups include, but are not limited to, benzimidazolyl,benzofuranyl, benzothiofuranyl, tetrahydrofuran, furanyl, furazanyl,imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl,indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl,isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl,isoxazolyl, methylenedioxyphenyl, morpholinyl, pyridinyl, pyridyl, andpyrimidinyl.

As used herein, the term “cycloalkyl” refers to a saturated orunsaturated nonaromatic hydrocarbon mono- or multi-ring (e.g., fused,bridged, or spiro rings) system having 3 to 10 carbon atoms (e.g.,C₃-C₆). Examples of cycloalkyl include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, cyclopentenyl, cyclohexenyl, and cycloheptenyl.

The term “heterocycloalkyl” refers to a saturated or unsaturatednonaromatic 3-8 membered monocyclic or bicyclic (fused, bridged, orspiro rings) having one or more heteroatoms (such as O, N, or S), unlessspecified otherwise. Examples of heterocycloalkyl groups include, butare not limited to, piperidinyl, piperazinyl, pyrrolidinyl, dioxanyl,tetrahydrofuranyl, isoindolinyl, indolinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, oxiranyl, azetidinyl, oxetanyl, thietanyl,1,2,3,6-tetrahydropyridinyl, tetrahydropyranyl, dihydropyranyl, pyranyl,morpholinyl, and tetrahydrothiopyranyl and the like.

As used herein, any recited moiety which includes, but is not limitedto, alkyl, alkenyl, alkynyl, alkoxy, carbocyclic ring, heterocyclicring, cycloalkyl, heterocycloalkyl, etc. can be optionally substituted.

The term “FXR modulator” refers to any compound that interacts with theFXR receptor. The interaction is not limited to a compound acting as anantagonist, agonist, partial agonist, or inverse agonist of the FXRreceptor. In one embodiment, the compound of the invention acts as anantagonist of the FXR receptor. In another aspect, the compound of theinvention acts as an agonist of the FXR receptor. In another aspect, thecompound of the invention acts as a partial agonist of the FXR receptor.In another aspect, the compound of the invention acts as an inverseagonist of the FXR receptor.

“Solvate”, as used herein, refers to a solvent addition form of acompound of the invention that contains either stoichiometric ornon-stoichiometric amounts of solvent. Some compounds have a tendency totrap a fixed molar ratio of solvent molecules in the crystalline solidstate, thus forming a solvate. If the solvent is water, the solvateformed is a hydrate, and when the solvent is alcohol, the solvate formedis an alcoholate. Hydrates are formed by the combination of one or moremolecules of water with one of the substances in which the water retainsits molecular state as H₂O, such combination being able to form one ormore hydrate.

As used herein, the term “amino acid conjugates” refers to conjugates ofa compound of the invention with any suitable amino acid. Taurine(—NH(CH₂)₂SO₃H), glycine (—NHCH₂CO₂H), and sarcosine (—N(CH₃)CH₂CO₂H)are examples of amino acid conjugates. Suitable amino acid conjugates ofthe compounds have the added advantage of enhanced integrity in bile orintestinal fluids. Suitable amino acids are not limited to taurine,glycine, and sarcosine.

As defined herein, the term “metabolite” refers to glucuronidated andsulphated derivatives of the compounds described herein, wherein one ormore glucuronic acid or sulphate moieties are linked to compound of theinvention. Glucuronic acid moieties may be linked to the compoundsthrough glycosidic bonds with the hydroxyl groups of the compounds(e.g., 3-hydroxyl, 7-hydroxyl, 11-hydroxyl, and/or the hydroxyl of theR⁷ group). Sulphated derivatives of the compounds may be formed throughsulphation of the hydroxyl groups (e.g., 3-hydroxyl, 7-hydroxyl,11-hydroxyl, and/or the hydroxyl of the R⁷ group). Examples ofmetabolites include, but are not limited to, 3-O-glucuronide,7-O-glucuronide, 11-O-glucuronide, 3-O-7-O-diglucuronide,3-O-11-O-triglucuronide, 7-O-11-O-triglucuronide, and3-O-7-O-11-O-triglucuronide, of the compounds described herein, and3-sulphate, 7-sulphate, 11-sulphate, 3,7-bisulphate, 3,11-bisulphate,7,11-bisulphate, and 3,7,11-trisulphate, of the compounds describedherein.

As used herein, “pharmaceutically acceptable salts” refer to derivativesof a compound of the invention wherein the parent compound is modifiedby forming acid or base salts thereof. Examples of pharmaceuticallyacceptable salts include, but are not limited to, mineral or organicacid salts of basic residues such as amines; alkali or organic salts ofacidic residues such as carboxylic acids; and the like. Thepharmaceutically acceptable salts include the conventional non-toxicsalts or the quaternary ammonium salts of the parent compound formed,for example, from non-toxic inorganic or organic acids. For example,such conventional non-toxic salts include, but are not limited to, thosederived from inorganic and organic acids selected from 2-acetoxybenzoic,2-hydroxyethane sulfonic, acetic, ascorbic, benzene sulfonic, benzoic,bicarbonic, carbonic, citric, edetic, ethane disulfonic, fumaric,glucoheptonic, gluconic, glutamic, glycolic, glycollyarsanilic,hexylresorcinic, hydrabamic, hydrobromic, hydrochloric, hydroiodide,hydroxymaleic, hydroxynaphthoic, isethionic, lactic, lactobionic, laurylsulfonic, maleic, malic, mandelic, methane sulfonic, napsylic, nitric,oxalic, pamoic, pantothenic, phenylacetic, phosphoric, polygalacturonic,propionic, salicylic, stearic, subacetic, succinic, sulphamic,sulphanilic, sulphuric, tannic, tartaric, and toluene sulphonic.

The phrase “pharmaceutically acceptable carrier” is art-recognized, andincludes, for example, pharmaceutically acceptable materials,compositions or vehicles, such as a liquid or solid filler, diluent,excipient, solvent or encapsulating material, involved in carrying ortransporting any subject composition from one organ, or portion of thebody, to another organ, or portion of the body. Each carrier is“acceptable” in the sense of being compatible with the other ingredientsof a subject composition and not injurious to the patient. In certainembodiments, a pharmaceutically acceptable carrier is non-pyrogenic.Some examples of materials which may serve as pharmaceuticallyacceptable carriers include: (1) sugars, such as lactose, glucose andsucrose; (2) starches, such as corn starch and potato starch; (3)cellulose, and its derivatives, such as sodium carboxymethyl cellulose,ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5)malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter andsuppository waxes; (9) oils, such as peanut oil, cottonseed oil,sunflower oil, sesame oil, olive oil, corn oil and soybean oil; (10)glycols, such as propylene glycol; (11) polyols, such as glycerin,sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyloleate and ethyl laurate; (13) agar; (14) buffering agents, such asmagnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16)pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19)ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxiccompatible substances employed in pharmaceutical formulations.

A “composition” or “pharmaceutical composition” is a formulationcontaining a compound of the invention or a salt, solvate, or amino acidconjugate thereof In one embodiment, the pharmaceutical composition isin bulk or in unit dosage form. The unit dosage form is any of a varietyof forms, including, for example, a capsule, an IV bag, a tablet, asingle pump on an aerosol inhaler, or a vial. The quantity of activeingredient (e.g., a formulation of a compound of the invention or saltsthereof) in a unit dose of composition is an effective amount and isvaried according to the particular treatment involved. One skilled inthe art will appreciate that it may be necessary to make routinevariations to the dosage depending on the age and condition of thepatient. The dosage will also depend on the route of administration. Avariety of routes are contemplated, including oral, ocular, ophthalmic,pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous,intramuscular, intraperitoneal, intranasal, and the like. Dosage formsfor the topical or transdermal administration of a compound of thisapplication include powders, sprays, ointments, pastes, creams, lotions,gels, solutions, patches and inhalants. In another embodiment, theactive compound is mixed under sterile conditions with apharmaceutically acceptable carrier, and with any preservatives,buffers, or propellants that are required.

The term “treating”, as used herein, refers to relieving, lessening,reducing, eliminating, modulating, or ameliorating, i.e., causingregression of the disease state or condition.

The term “preventing”, as used herein, refers to completely or almostcompletely stop a disease state or condition, from occurring in apatient or subject, especially when the patient or subject ispredisposed to such or at risk of contracting a disease state orcondition. Preventing can also include inhibiting, i.e., arresting thedevelopment, of a disease state or condition, and relieving orameliorating, i.e., causing regression of the disease state orcondition, for example when the disease state or condition may alreadybe present.

The phrase “reducing the risk of”, as used herein, refers to loweringthe likelihood or probability of a central nervous system disease,inflammatory disease and/or metabolic disease from occurring in apatient, especially when the subject is predisposed to such occurrence.

“Combination therapy” (or “co-therapy”) refers to the administration ofa compound of the invention and at least a second agent as part of aspecific treatment regimen intended to provide the beneficial effectfrom the co-action of these therapeutic agents (i.e., the compound ofthe invention and at least a second agent). The beneficial effect of thecombination includes, but is not limited to, pharmacokinetic orpharmacodynamic co-action resulting from the combination of therapeuticagents. Administration of these therapeutic agents in combinationtypically is carried out over a defined time period (usually minutes,hours, days or weeks depending upon the combination selected).“Combination therapy” may, but generally is not, intended to encompassthe administration of two or more of these therapeutic agents as part ofseparate monotherapy regimens that incidentally and arbitrarily resultin the combinations of the present application. “Combination therapy” isintended to embrace administration of these therapeutic agents in asequential manner, that is, wherein each therapeutic agent isadministered at a different time, as well as administration of thesetherapeutic agents, or at least two of the therapeutic agents, in asubstantially simultaneous manner. Substantially simultaneousadministration can be accomplished, for example, by administering to thesubject a single capsule having a fixed ratio of each therapeutic agentor in multiple, single capsules for each of the therapeutic agents.Sequential or substantially simultaneous administration of eachtherapeutic agent can be effected by any appropriate route including,but not limited to, oral routes, intravenous routes, intramuscularroutes, and direct absorption through mucous membrane tissues. Thetherapeutic agents can be administered by the same route or by differentroutes. For example, a first therapeutic agent of the combinationselected may be administered by intravenous injection while the othertherapeutic agents of the combination may be administered orally.Alternatively, for example, all therapeutic agents may be administeredorally or all therapeutic agents may be administered by intravenousinjection. The sequence in which the therapeutic agents are administeredis not narrowly critical.

“Combination therapy” also embraces the administration of thetherapeutic agents as described above in further combination with otherbiologically active ingredients and non-drug therapies (e.g., surgery ormechanical treatments). Where the combination therapy further comprisesa non-drug treatment, the non-drug treatment may be conducted at anysuitable time so long as a beneficial effect from the co-action of thecombination of the therapeutic agents and non-drug treatment isachieved. For example, in appropriate cases, the beneficial effect isstill achieved when the non-drug treatment is temporally removed fromthe administration of the therapeutic agents, perhaps by days or evenweeks.

An “effective amount” of a compound of the invention, or a combinationof compounds is an amount (quantity or concentration) of compound orcompounds. In one embodiment, when a therapeutically effective amount ofa compound is administered to a subject in need of treatment symptomsarising from the disease are ameliorated immediately or afteradministration of the compound one or more times. The amount of thecompound to be administered to a subject will depend on the particulardisorder, the mode of administration, co-administered compounds, if any,and the characteristics of the subject, such as general health, otherdiseases, age, sex, genotype, body weight, and tolerance to drugs. Theskilled artisan will be able to determine appropriate dosages dependingon these and other factors.

The term “prophylactically effective amount” means an amount (quantityor concentration) of a compound of the present invention, or acombination of compounds, that is administered to prevent or reduce therisk of a disease—in other words, an amount needed to provide apreventative or prophylactic effect. The amount of the present compoundto be administered to a subject will depend on the particular disorder,the mode of administration, co-administered compounds, if any, and thecharacteristics of the subject, such as general health, other diseases,age, sex, genotype, body weight, and tolerance to drugs. The skilledartisan will be able to determine appropriate dosages depending on theseand other factors. A “subject” includes mammals, e.g., humans, companionanimals (e.g., dogs, cats, birds, and the like), farm animals (e.g.,cows, sheep, pigs, horses, and the like), and laboratory animals (e.g.,rats, mice, guinea pigs, and the like). Typically, the subject is human.

As used herein, farnesoid X receptor or FXR refers to all mammalianforms of such receptor including, for example, alternative spliceisoforms and naturally occurring isoforms (see, e.g., Huber et al., Gene290:35-43 (2002)). Representative FXR species include, withoutlimitation rat FXR (GenBank Accession No. NM 021745), mouse FXR (GenBankAccession No. NM 009108), and human FXR (GenBank Accession No. NM005123).

Compounds of the Invention

In one aspect, the present disclosure provides a compound of formula I:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein:

R¹ is OH, alkoxy, halogen, or oxo;

R² and R³ are each independently H, OH, halogen, or alkyl optionallysubstituted with one or more halogen or OH, or R² and R³ taken togetherwith the carbon atom to which they are attached form a carbonyl;

R⁴ is H, halogen, alkyl optionally substituted with one or more halogenor OH, alkenyl, or alkynyl;

R⁵ and R⁶ are each independently H, OH, OSO₃H, OCOCH₃, OPO₃H₂, orhalogen, or R⁵ and R⁶ taken together with the carbon atom to which theyare attached form a carbonyl;

R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂, SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, C(O)NHOH,tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or 2,4-difluoro-3-hydroxyphenyl;

R⁸, R⁹, and R¹⁰ are each independently H, OH, halogen, or alkyloptionally substituted with one or more halogen or OH, or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2heteroatoms selected from N, O, and S, or R⁹ and R¹⁰ taken together withthe carbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S;

m is 0, 1, or 2;

n is 0 or 1;

p is 0 or 1; and

is a single or double bond, provided that when each

is a single bond, the sum of m, n, and p is 2, R¹ is OH, and R⁸, R⁹, andR¹⁰ are each H, then R⁷ is not CO₂H.

In one of the embodiments, the present disclosure provides a compound offormula Ia:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one of the embodiments, the present disclosure provides a compound offormula Ib or Ic:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R¹ is OH, alkoxy, or oxo.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R¹ is OH or alkoxy.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R¹ is halogen.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R¹ is OH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R¹ is alkoxy.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R³ is H.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R³ is OH or halogen.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R³ is C₁-C₆ alkyl optionally substitutedwith one or more halogen or OH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R⁶ is OH or H.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Ic, wherein R⁶ is OSO₃H, OCOCH₃, or OPO₃H₂.

In one of the embodiments, the present disclosure a compound of formulaId:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one of the embodiments, the present disclosure a compound is offormula Ie:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂, SO₂NH₂,OPO₃H₂, PO₃H₂, CO₂H, or C(O)NHOH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OH, OSO₃H, OSO₂NH₂, OPO₃H₂, or CO₂H.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is CO₂H.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OSO₃H.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is SO₃H.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OSO₂NH₂ or SO₂NH₂.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OPO₃H₂, PO₃H₂, or C(O)NHOH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is tetrazolyl, oxadiazolyl,thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl, 5-oxo-1,2,4-thiadiazolyl,oxazolidine-dionyl, thiazolidine-dionyl, 3-hydroxyisoxazolyl,3-hydroxyisothiazolyl, or 2,4-difluoro-3-hydroxyphenyl.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OH, OSO₃H, OSO₂NH₂, OPO₃H₂, CO₂H,tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or2,4-difluoro-3-hydroxyphenyl.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R² is OH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R² is H or halogen.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R² is C₁-C₆ alkyl optionally substitutedwith one or more halogen or OH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁵ is OH or H.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁵ is OSO₃H, OCOCH₃, or OPO₃H₂.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁵ and R⁶ taken together with the carbonatom to which they are attached form a carbonyl.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁴ is H or halogen.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁴ is C₁-C₆ alkyl optionally substitutedwith one or more halogen or OH.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁴ is C₂-C₆ alkenyl or C₂-C₆ alkynyl.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁴ is methyl, ethyl, or propyl.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein m is 0.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein m is 1.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein m is 2.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein n is 1.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein p is 0.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁴ is in the a-position.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R¹ is in the β-position.

In one of the embodiments, the present disclosure provides compounds offormula I, wherein the compound is selected from:

In one of the embodiments, the present disclosure provides salts ofcompounds of formula I and Ia-Id.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OSO₃ ⁻.

In one of the embodiments, the present disclosure provides compounds offormula I and Ia-Id, wherein R⁷ is OSO₃ ⁻Na⁺.

In one of the embodiments, the present disclosure provides a compound offormula I

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein:

R¹ is alkoxy or oxo;

R² and R³ are each independently H, OH, halogen, or alkyl optionallysubstituted with one or more halogen or OH, or R² and R³ taken togetherwith the carbon atom to which they are attached form a carbonyl;

R⁴ is H, halogen, alkyl optionally substituted with one or more halogenor OH, alkenyl, or alkynyl;

R⁵ and R⁶ are each independently H, OH, OSO₃H, OCOCH₃, OPO₃H₂, orhalogen, or R⁵ and R⁶ taken together with the carbon atom to which theyare attached form a carbonyl;

R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂, SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, C(O)NHOH,tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or2,4-difluoro-3-hydroxyphenyl;

R⁸, R⁹, and R¹⁰ are each independently H, OH, halogen, or alkyloptionally substituted with one or more halogen or OH, or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2heteroatoms selected from N, O, and S, or R⁹ and R¹⁰ taken together withthe carbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S;

m is 0, 1, or 2;

n is 0 or 1;

p is 0 or 1; and

is a single or double bond.

In one of the embodiments, the present disclosure provides a compound offormula I:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein:

R¹ is OH, alkoxy, halogen, or oxo;

R² and R³ are each independently H, OH, halogen, or alkyl optionallysubstituted with one or more halogen or OH, or R² and R³ taken togetherwith the carbon atom to which they are attached form a carbonyl;

R⁴ is H, halogen, alkyl optionally substituted with one or more halogenor OH, alkenyl, or alkynyl;

R⁵ and R⁶ are each independently H, OH, OSO₃H, OCOCH₃, OPO₃H₂, orhalogen, or R⁵ and R⁶ taken together with the carbon atom to which theyare attached form a carbonyl;

R⁷ is OSO₃H, OSO₂NH₂, OPO₃H₂, C(O)NHOH, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or2,4-difluoro-3-hydroxyphenyl;

R⁸, R⁹, and R¹⁰ are each independently H, OH, halogen, or alkyloptionally substituted with one or more halogen or OH, or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2heteroatoms selected from N, O, and S, or R⁹ and R¹⁰ taken together withthe carbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S;

m is 0, 1, or 2;

n is 0 or 1;

p is 0 or 1; and

is a single or double bond.

In one of the embodiments, the present disclosure provides a compound offormula I:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein:

R¹ is OH, alkoxy, halogen, or oxo;

R² is H, OH, halogen, or alkyl optionally substituted with one or morehalogen or OH, or R² and R³ taken together with the carbon atom to whichthey are attached form a carbonyl;

R³ is OH, halogen, or alkyl optionally substituted with one or morehalogen or OH, or R² and R³ taken together with the carbon atom to whichthey are attached form a carbonyl;

R⁴ is H, halogen, alkyl optionally substituted with one or more halogenor OH, alkenyl, or alkynyl;

R⁵ and R⁶ are each independently H, OH, OSO₃H, OCOCH₃, OPO₃H₂, orhalogen, or R⁵ and R⁶ taken together with the carbon atom to which theyare attached form a carbonyl;

R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂, SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, C(O)NHOH,tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or2,4-difluoro-3-hydroxyphenyl;

R⁸, R⁹, and R¹⁰ are each independently H, OH, halogen, or alkyloptionally substituted with one or more halogen or OH, or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2heteroatoms selected from N, O, and S, or R⁹ and R¹⁰ taken together withthe carbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S;

m is 0, 1, or 2;

n is 0 or 1;

p is 0 or 1; and

is a single or double bond.

In one of the embodiments, the present disclosure a compound of formulaI:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein:

R¹ is OH, alkoxy, halogen, or oxo;

R² and R³ are each independently H, OH, halogen, or alkyl optionallysubstituted with one or more halogen or OH, or R² and R³ taken togetherwith the carbon atom to which they are attached form a carbonyl;

R⁴ is H, halogen, alkyl optionally substituted with one or more halogenor OH, alkenyl, or alkynyl;

R⁵ is OH, OSO₃H, OCOCH₃, OPO₃H₂, or halogen;

R⁶ is H, OH, OSO₃H, OCOCH₃, OPO₃H₂, or halogen, or R⁵ and R⁶ takentogether with the carbon atom to which they are attached form acarbonyl;

R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂, SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, C(O)NHOH,tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or2,4-difluoro-3-hydroxyphenyl;

R⁸, R⁹, and R¹⁰ are each independently H, OH, halogen, or alkyloptionally substituted with one or more halogen or OH, or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2heteroatoms selected from N, O, and S, or R⁹ and R¹⁰ taken together withthe carbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S;

m is 0, 1, or 2;

n is 0 or 1;

p is 0 or 1; and

is a single or double bond.

In one of the embodiments, the present disclosure provides a compound offormula I:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein:

R¹ is OH, alkoxy, halogen, or oxo;

R² and R³ are each independently H, OH, halogen, or alkyl optionallysubstituted with one or more halogen or OH, or R² and R³ taken togetherwith the carbon atom to which they are attached form a carbonyl;

R⁴ is H, halogen, alkyl optionally substituted with one or more halogenor OH, alkenyl, or alkynyl;

R⁵ is H, OH, OSO₃H, OCOCH₃, OPO₃H₂, or halogen;

R⁶ is OH, OSO₃H, OCOCH₃, OPO₃H₂, or halogen, or R⁵ and R⁶ taken togetherwith the carbon atom to which they are attached form a carbonyl;

R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂, SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, C(O)NHOH,tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or2,4-difluoro-3-hydroxyphenyl;

R⁸, R⁹, and R¹⁰ are each independently H, OH, halogen, or alkyloptionally substituted with one or more halogen or OH, or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2heteroatoms selected from N, O, and S, or R⁹ and R¹⁰ taken together withthe carbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S;

m is 0, 1, or 2;

n is 0 or 1;

p is 0 or 1; and

is a single or double bond.

In one of the embodiments, the present disclosure provides a compound offormula I, wherein R₇ is OH.

In one of the embodiments, the present disclosure provides a compound offormula I, wherein the compound is selected from:

In one embodiment, the compound is of formula II:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In another embodiment, the compound is of formula III:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In yet another embodiment, the compound is of formula IV:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one embodiment, the compound is of formula V:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In another embodiment, the compound is of formula VI:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In another embodiment, the compound is of formula VII:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH. In anotherembodiment, the present disclosure relates to a compound of formula I,II, III, IV, V, VI, or VI, wherein R¹ is halogen. In another embodiment,R¹ is fluoro. In another embodiment, the present disclosure relates to acompound of formula I, II, III, IV, V, VI, or VII, wherein R¹ is C₁-C₆alkoxy. In one embodiment, the present disclosure relates to a compoundof formula I, II, or III, wherein R¹ is oxo.

In another embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R² is OH. In oneembodiment, the present disclosure relates to a compound of formula I,II, III, IV, V, VI, or VII, wherein R² is H or halogen. In anotherembodiment, the present disclosure relates to a compound of formula I,II, III, IV, V, VI, or VII, wherein R² is C₁-C₆ alkyl optionallysubstituted with one or more halogen or OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH and R² is OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH and R² is H orhalogen.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH and R² is C₁-C₆alkyl optionally substituted with one or more halogen or OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R³ is H. In oneembodiment, the present disclosure relates to a compound of formula I,II, III, IV, V, VI, or VII wherein R³ is OH or halogen. In anotherembodiment, the present disclosure relates to a compound of formula I,II, III, IV, V, VI, or VII, wherein R³ is C₁-C₆ alkyl optionallysubstituted with one or more halogen or OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, andR³ is H.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, andR³ is OH or halogen.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, andR³ is C₁-C₆ alkyl optionally substituted with one or more halogen or OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R² and R³ taken togetherwith the carbon atom to which they are attached form a carbonyl.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁴ is H or halogen. Inone embodiment, the present disclosure relates to a compound of formulaI, II, III, IV, V, VI, or VII, wherein R⁴ is C₁-C₆ alkyl optionallysubstituted with one or more halogen or OH. In one embodiment, R⁴ isC₂-C₆ alkenyl or alkynyl. In one embodiment, R⁴ is methyl, ethyl, orpropyl. In one embodiment, R⁴ is ethyl.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, R³ isH, and R⁴ is methyl, ethyl, or propyl.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, R³ isH, and R⁴ is C₁-C₆ alkyl optionally substituted with one or more halogenor OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, R³ isH, and R⁴ is C₂-C₆ alkenyl or alkynyl.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁵ is OH or H. In anotherembodiment, R⁵ is OSO₃H, OCOCH₃, or OPO₃H₂. In another embodiment, R⁵ ishalogen.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, R³ isH, R⁴ is methyl, ethyl, or propyl, and R⁵ is OH or H.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R¹ is OH, R² is OH, R³ isH, R⁴ is C₁-C₆ alkyl optionally substituted with one or more halogen orOH, and R⁵ is OH or H.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁶ is OH or H. In anotherembodiment, the present disclosure relates to a compound of formula I,II, III, IV, V, VI, or VII, wherein R⁶ is OSO₃H, OCOCH₃, or OPO₃H₂. Inanother embodiment, R⁶ is halogen.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁵ and R⁶ taken togetherwith the carbon atom to which they are attached form a carbonyl.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁷ is OH. In oneembodiment, R⁷ is CO₂H. In one embodiment, the present disclosurerelates to a compound of formula I, II, III, IV, V, VI, or VII, whereinR⁷ is OSO₃H. In a separate embodiment, the present disclosure relates toa compound of formula I, II, III, IV, V, VI, or VII, wherein R⁷ is SO₃H.In another embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁷ is OSO₂NH₂ or SO₂NH₂.In a separate embodiment, the present disclosure relates to a compoundof formula I, II, III, IV, V, VI, or VII, wherein R⁷ is OPO₃H₂, PO₃H₂,or C(O)NHOH.

In a separate embodiment, the present disclosure relates to a compoundof formula I, II, III, IV, V, VI, or VII, wherein R⁷ is tetrazolyl,oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl,5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl,3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, or2,4-difluoro-3-hydroxyphenyl. In a separate embodiment, the presentdisclosure relates to a compound of formula I, II, III, IV, V, VI, orVII, wherein R⁷ is tetrazolyl. In a separate embodiment, the presentdisclosure relates to a compound of formula I, II, III, IV, V, VI, orVII, wherein R⁷ is oxadiazolyl. In a separate embodiment, the presentdisclosure relates to a compound of formula I, II, III, IV, V, VI, orVII, wherein R⁷ is thiadiazolyl. In a separate embodiment, the presentdisclosure relates to a compound of formula I, II, III, IV, V, VI, orVII, wherein R⁷ is 5-oxo-1,2,4-oxadiazolyl. In one embodiment, thepresent disclosure relates to a compound of formula I, II, III, IV, V,VI, or VII, wherein R⁷ is oxazolidine-dionyl. In one embodiment, thepresent disclosure relates to a compound of formula I, II, III, IV, V,VI, or VII, wherein R⁷ is thiazolidine-dionyl. In one embodiment, thepresent disclosure relates to a compound of formula I, II, III, IV, V,VI, or VII, wherein R⁷ is 2,4-difluoro-3-hydroxyphenyl.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁸ is H. In oneembodiment, R⁸ is independently is H or OH. In one embodiment, R⁸ isindependently H or halogen. In one embodiment, R⁸ is independently H oralkyl. In one embodiment, R⁸ is independently H or alkyl substitutedwith one or more halogen or OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁹ is H. In oneembodiment, R⁹ is H or OH. In one embodiment, R⁹ is H or halogen. In oneembodiment, R⁹ is H or alkyl. In one embodiment, R⁹ is H or alkylsubstituted with one or more halogen or OH.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein R⁸ and R⁹ are alkyl andtaken together with the carbons to which they are attached form a ringof size 3, 4, 5, or 6 atoms.

In one embodiment, the present disclosure relates to a compound offormula I, VI, or VII, wherein R¹⁰ is H. In one embodiment, R¹⁰ is H orOH. In one embodiment, R¹⁰ is H or halogen. In one embodiment, R¹⁰ is Hor alkyl. In one embodiment, R¹⁰ is H or alkyl substituted with one ormore halogen or OH.

In one embodiment, the present disclosure relates to a compound offormula I, VI, or VII, wherein R⁹ and R¹⁰ are alkyl and taken togetherwith the carbons to which they are attached form a ring of size 3, 4, 5,or 6 atoms.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein m is 0. In oneembodiment, the present disclosure relates to a compound of formula I,II, III, IV, V, VI, or VII, wherein m is 1. In one embodiment, thepresent disclosure relates to a compound of formula I, II, III, IV, V,VI, or VII, wherein m is 2.

In one embodiment, the present disclosure relates to a compound offormula I, II, III, IV, V, VI, or VII, wherein n is 0. In oneembodiment, n is 1.

In one embodiment, the present disclosure relates to a compound offormula I, VI, or VII, wherein p is 0. In one embodiment, p is 1.

In one embodiment, the present disclosure relates to a compound offormula I, wherein the compound is selected from:

In one of the embodiments, the present disclosure relates to a compoundof formula I, wherein the compound is selected from:

The compounds of the invention have asymmetric centers and can beisolated in optically active or racemic forms. It is well known in theart how to prepare optically active forms, such as by resolution ofracemic forms or by synthesis from optically active starting materials.Many geometric isomers of olefins, and the like can also be present inthe compounds described herein, and all such stable isomers arecontemplated in the present invention. Cis and trans geometric isomersof the compounds of the invention and can be isolated as a mixture ofisomers or as separate isomeric forms. All chiral, diastereomeric,racemic, and geometric isomeric forms of a structure are intended,unless specific stereochemistry or isomeric form is specificallyindicated. All processes used to prepare compounds of the presentinvention and intermediates made therein are considered to be part ofthe present invention. All tautomers of shown or described compounds arealso considered to be part of the present invention. Furthermore, theinvention also includes metabolites of the compounds described herein.

The invention also comprehends isotopically-labelled compounds of theinvention, or pharmaceutically acceptable salts, solvates, or amino acidconjugates thereof, which are identical to those recited in formulae ofthe application and following, but for the fact that one or more atomsare replaced by an atom having an atomic mass or mass number differentfrom the atomic mass or mass number most commonly found in nature.Examples of isotopes that can be incorporated into compounds of theinvention, or pharmaceutically acceptable salts, solvates, or amino acidconjugates thereof include isotopes of hydrogen, carbon, nitrogen,fluorine, such as ³H, ¹¹C, ¹⁴C and ¹⁸F.

Tritiated, i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes may be used fortheir ease of preparation and detectability. Further, substitution withheavier isotopes such as deuterium, i.e., ²H, can afford certaintherapeutic advantages resulting from greater metabolic stability, forexample increased in vivo half-life or reduced dosage requirements and,hence, may be used in some circumstances. Isotopically labelledcompounds of the invention, or pharmaceutically acceptable salts,solvates, or amino acid conjugates thereof can generally be prepared bycarrying out the procedures disclosed in the Schemes and/or in theExamples, by substituting a readily available isotopically labelledreagent for a non-isotopically labelled reagent. However, one skilled inthe art will recognize that not all isotopes can be included bysubstitution of the non-isotopically labelled reagent. In oneembodiment, compounds of the invention, or pharmaceutically acceptablesalts, solvates, or amino acid conjugates thereof are not isotopicallylabelled. In one embodiment, deuterated compounds of the invention areuseful for bioanalytical assays. In another embodiment, compounds of theinvention, or pharmaceutically acceptable salts, solvates, or amino acidconjugates thereof are radiolabelled.

Pharmaceutical Compositions

A “pharmaceutical composition” is a formulation containing one or morecompounds of the invention in a form suitable for administration to asubject. In one embodiment, the pharmaceutical composition is in bulk orin unit dosage form. It can be advantageous to formulate compositions indosage unit form for ease of administration and uniformity of dosage.Dosage unit form as used herein refers to physically discrete unitssuited as unitary dosages for the subject to be treated; each unitcontaining a predetermined quantity of active reagent calculated toproduce the desired therapeutic effect in association with the requiredpharmaceutical carrier. The specification for the dosage unit forms aredictated by and directly dependent on the unique characteristics of theactive reagent and the particular therapeutic effect to be achieved, andthe limitations inherent in the art of compounding such an active agentfor the treatment of individuals.

Possible formulations include those suitable for oral, sublingual,buccal, parenteral (e.g., subcutaneous, intramuscular, or intravenous),rectal, topical including transdermal, intranasal and inhalationadministration. Most suitable means of administration for a particularpatient will depend on the nature and severity of the disease beingtreated or the nature of the therapy being used and on the nature of theactive compound, but where possible, oral administration may be used forthe prevention and treatment of FXR mediated diseases and conditions.Formulations suitable for oral administration may be provided asdiscrete units, such as tablets, capsules, cachets, lozenges, eachcontaining a predetermined amount of the active compound; as powders orgranules; as solutions or suspensions in aqueous or non-aqueous liquids;or as oil-in- water or water-in-oil emulsions. Formulations suitable forsublingual or buccal administration include lozenges comprising theactive compound and, typically a flavored base, such as sugar and acaciaor tragacanth and pastilles comprising the active compound in an inertbase, such as gelatin and glycerin or sucrose acacia.

Formulations suitable for parenteral administration typically comprisesterile aqueous solutions containing a predetermined concentration ofthe active compound; the solution may be isotonic with the blood of theintended recipient. Additional formulations suitable for parenteraladministration include formulations containing physiologically suitableco-solvents and/or complexing agents such as surfactants andcyclodextrins. Oil-in-water emulsions are also suitable formulations forparenteral formulations. Although such solutions may be administeredintravenously, they may also be administered by subcutaneous orintramuscular injection.

Formulations suitable for rectal administration may be provided asunit-dose suppositories comprising the active ingredient in one or moresolid carriers forming the suppository base, for example, cocoa butter.

Formulations suitable for topical or intranasal application includeointments, creams, lotions, pastes, gels, sprays, aerosols, and oils.Suitable carriers for such formulations include petroleum jelly,lanolin, polyethyleneglycols, alcohols, and combinations thereof.

Formulations of the invention may be prepared by any suitable method,typically by uniformly and intimately admixing the active compound withliquids or finely divided solid carriers or both, in the requiredproportions and then, if necessary, shaping the resulting mixture intothe desired shape.

For example, a tablet may be prepared by compressing an intimate mixturecomprising a powder or granules of the active ingredient and one or moreoptional ingredients, such as a binder, lubricant, inert diluent, orsurface active dispersing agent, or by moulding an intimate mixture ofpowdered active ingredient and inert liquid diluent. Suitableformulations for administration by inhalation include fine particledusts or mists which may be generated by means of various types ofmetered dose pressurized aerosols, nebulizers, or insufflators.

For pulmonary administration via the mouth, the particle size of thepowder or droplets is typically in the range of 0.5-10 or may be about1-5 to ensure delivery into the bronchial tree. For nasaladministration, a particle size in the range of 10-500 μm may be used toensure retention in the nasal cavity.

Metered dose inhalers are pressurized aerosol dispensers, typicallycontaining a suspension or solution formulation of the active ingredientin a liquefied propellant. During use, these devices discharge theformulation through a valve adapted to deliver a metered volume,typically from 10 to 150 to produce a fine particle spray containing theactive ingredient. Suitable propellants include certainchlorofluorocarbon compounds, for example, dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, and mixtures thereof.The formulation may additionally contain one or more co-solvents, forexample, ethanol surfactants, such as oleic acid or sorbitan trioleate,anti-oxidants and suitable flavoring agents.

Nebulizers are commercially available devices that transform solutionsor suspensions of the active ingredient into a therapeutic aerosol misteither by means of acceleration of a compressed gas typically air oroxygen, through a narrow venturi orifice, or by means of ultrasonicagitation. Suitable formulations for use in nebulizers consist of theactive ingredient in a liquid carrier and comprise up to 40% w/w of theformulation, preferably less than 20% w/w. The carrier is typicallywater or a dilute aqueous alcoholic solution, preferably made isotonicwith body fluids by the addition of, for example, sodium chloride.Optional additives include preservatives if the formulation is notprepared sterile, for example, methyl hydroxy-benzoate, anti-oxidants,flavouring agents, volatile oils, buffering agents, and surfactants.

Suitable formulations for administration by insufflation include finelycomminuted powders which may be delivered by means of an insufflator ortaken into the nasal cavity in the manner of a snuff. In theinsufflator, the powder is contained in capsules or cartridges,typically made of gelatin or plastic, which are either pierced or openedin situ and the powder delivered by air drawn through the device uponinhalation or by means of a manually-operated pump. The powder employedin the insufflator consists either solely of the active ingredient or ofa powder blend comprising the active ingredient, a suitable powderdiluent, such as lactose, and an optional surfactant. The activeingredient typically comprises from 0.1 to 100% w/w of the formulation.

In a further embodiment, the present invention provides a pharmaceuticalcomposition comprising, as active ingredient, a compound of theinvention together, and/or in admixture, with at least onepharmaceutical carrier or diluent. These pharmaceutical compositions maybe used in the prevention or treatment of the foregoing diseases orconditions.

The carrier is pharmaceutically acceptable and must be compatible with,i.e. not have a deleterious effect upon, the other ingredients in thecomposition. The carrier may be a solid or liquid and is preferablyformulated as a unit dose formulation, for example, a tablet which maycontain from 0.05 to 95% by weight of the active ingredient. If desired,other physiologically active ingredients may also be incorporated in thepharmaceutical compositions of the invention.

In addition to the ingredients specifically mentioned above, theformulations of the present invention may include other agents known tothose skilled in the art of pharmacy, having regard for the type offormulation in issue. For example, formulations suitable for oraladministration may include flavouring agents and formulations suitablefor intranasal administration may include perfumes.

In one of the embodiments, the present disclosure provides apharmaceutical composition comprising the compounds of formula I andIa-Id and a pharmaceutically acceptable carrier or excipient.

Methods of Treatment

The compounds of the invention are useful for therapy in subjects suchas mammals, including humans. In particular, the compounds of theinvention are useful in a method of treating or preventing a disease orcondition in a subject comprising administering to the subject in needthereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof. In one embodiment, the disease or condition is FXR-mediated(e.g., FXR plays a role in the initiation or progress of the disease orcondition). In one embodiment, the disease or condition is mediated bydecreased FXR activity. In one embodiment, the disease or condition isselected from cardiovascular disease, chronic liver disease, lipiddisorder, gastrointestinal disease, renal disease, metabolic disease,cancer, and neurological disease.

In one embodiment, the invention relates to a method of treating orpreventing cardiovascular disease in a subject, comprising administeringto the subject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating cardiovascular disease. In one embodiment, cardiovasculardisease selected from atherosclerosis, arteriosclerosis, dyslipidemia,hypercholesteremia, hyperlipidemia, hyperlipoproteinemia, andhypertriglyceridemia.

The term “hyperlipidemia” refers to the presence of an abnormallyelevated level of lipids in the blood. Hyperlipidemia can appear in atleast three forms: (1) hypercholesterolemia, i.e., an elevatedcholesterol level; (2) hypertriglyceridemia, i.e., an elevatedtriglyceride level; and (3) combined hyperlipidemia, i.e., a combinationof hypercholesterolemia and hypertriglyceridemia.

The term “dyslipidemia” refers to abnormal levels of lipoproteins inblood plasma including both depressed and/or elevated levels oflipoproteins (e.g., elevated levels of LDL, VLDL and depressed levels ofHDL).

In one embodiment, the invention relates to a method selected fromreducing cholesterol levels or modulating cholesterol metabolism,catabolism, absorption of dietary cholesterol, and reverse cholesteroltransport in a subject, comprising administering to the subject in needthereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In another embodiment, the invention relates to a method of treating orpreventing a disease affecting cholesterol, triglyceride, or bile acidlevels in a subject, comprising administering to the subject in needthereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one embodiment, the invention relates to a method of loweringtriglycerides in a subject, comprising administering to the subject inneed thereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one embodiment, the invention relates to a method of treating orpreventing a disease state associated with an elevated cholesterol levelin a subject, comprising administering to the subject in need thereof aneffective amount of a compound of the invention or a pharmaceuticallyacceptable salt, solvate, or amino acid conjugate thereof. In oneembodiment, the invention relates to a method of treating a diseasestate associated with an elevated cholesterol level in a subject. In oneembodiment, the invention relates to a method of preventing a diseasestate associated with an elevated cholesterol level in a subject. In oneembodiment, the disease state is selected from coronary artery disease,angina pectoris, carotid artery disease, strokes, cerebralarteriosclerosis, and xanthoma.

In one embodiment, the invention relates to a method of treating orpreventing a lipid disorder in a subject, comprising administering tothe subject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating a lipid disorder. In one embodiment, the invention relatesto a method of preventing a lipid disorder.

Lipid disorders are the term for abnormalities of cholesterol andtriglycerides. Lipid abnormalities are associated with an increased riskfor vascular disease, and especially heart attacks and strokes.Abnormalities in lipid disorders are a combination of geneticpredisposition as well as the nature of dietary intake. Many lipiddisorders are associated with being overweight. Lipid disorders may alsobe associated with other diseases including diabetes, the metabolicsyndrome (sometimes called the insulin resistance syndrome), underactivethyroid or the result of certain medications (such as those used foranti-rejection regimens in people who have had transplants).

In one embodiment, the invention relates to a method of treating orpreventing one or more symptoms of disease affecting lipid metabolism(i.e., lipodystrophy) in a subject, comprising administering to thesubject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating one or more symptoms of a disease affecting lipidmetabolism. In one embodiment, the invention relates to a method ofpreventing one or more symptoms of a disease affecting lipid metabolism.

In one embodiment, the invention relates to a method of decreasing lipidaccumulation in a subject, comprising administering to the subject inneed thereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.

In one embodiment, the invention relates to a method of treating orpreventing chronic liver disease in a subject, comprising administeringto the subject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating chronic liver disease. In one embodiment, the inventionrelates to a method of preventing chronic liver disease. In oneembodiment, the chronic liver disease is selected from primary biliarycirrhosis (PBC), cerebrotendinous xanthomatosis (CTX), primarysclerosing cholangitis (PSC), drug induced cholestasis, intrahepaticcholestasis of pregnancy, parenteral nutrition associated cholestasis(PNAC), bacterial overgrowth or sepsis associated cholestasis,autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease,nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis(NASH), liver transplant associated graft versus host disease, livingdonor transplant liver regeneration, congenital hepatic fibrosis,choledocholithiasis, granulomatous liver disease, intra- or extrahepaticmalignancy,

Sjogren's syndrome, Sarcoidosis, Wilson's disease, Gaucher's disease,hemochromatosis, and alpha 1-antitrypsin deficiency.

In one embodiment, the invention relates to a method of treating orpreventing one or more symptoms of cholestasis, including complicationsof cholestasis in a subject, comprising administering to the subject inneed thereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof. In one embodiment, the invention relates to a method oftreating one or more symptoms of cholestasis. In one embodiment, theinvention relates to preventing one or more symptoms of cholestasis.

Cholestasis is typically caused by factors within the liver(intrahepatic) or outside the liver (extrahepatic) and leads to theaccumulation of bile salts, bile pigment bilirubin, and lipids in theblood stream instead of being eliminated normally. Intrahepaticcholestasis is characterized by widespread blockage of small ducts or bydisorders, such as hepatitis, that impair the body's ability toeliminate bile. Intrahepatic cholestasis may also be caused by alcoholicliver disease, primary biliary cirrhosis, cancer that has spread(metastasized) from another part of the body, primary sclerosingcholangitis, gallstones, biliary colic, and acute cholecystitis. It canalso occur as a complication of surgery, serious injury, cysticfibrosis, infection, or intravenous feeding or be drug induced.Cholestasis may also occur as a complication of pregnancy and oftendevelops during the second and third trimesters. Extrahepaticcholestasis is most often caused by choledocholithiasis (Bile DuctStones), benign biliary strictures (non-cancerous narrowing of thecommon duct), cholangiocarcinoma (ductal carcinoma), and pancreaticcarcinoma. Extrahepatic cholestasis can occur as a side effect of manymedications.

A compound of the invention may be used for treating or preventing oneor more symptoms of intrahepatic or extrahepatic cholestasis, includingwithout limitation, biliary atresia, obstetric cholestasis, neonatalcholestasis, drug induced cholestasis, cholestasis arising fromHepatitis C infection, chronic cholestatic liver disease such as primarybiliary cirrhosis (PBC), and primary sclerosing cholangitis (PSC).

In one embodiment, the invention relates to a method of enhancing liverregeneration in a subject, comprising administering to the subject inneed thereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof. In one embodiment, the method is enhancing liver regenerationfor liver transplantation.

In one embodiment, the invention relates to a method of treating orpreventing fibrosis in a subject, comprising administering to thesubject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating fibrosis. In one embodiment, the invention relates to amethod of preventing fibrosis.

Accordingly, as used herein, the term fibrosis refers to all recognizedfibrotic disorders, including fibrosis due to pathological conditions ordiseases, fibrosis due to physical trauma (“traumatic fibrosis”),fibrosis due to radiation damage, and fibrosis due to exposure tochemotherapeutics. As used herein, the term “organ fibrosis” includesbut is not limited to liver fibrosis, fibrosis of the kidneys, fibrosisof lung, and fibrosis of the intestine. “Traumatic fibrosis” includesbut is not limited to fibrosis secondary to surgery (surgical scarring),accidental physical trauma, burns, and hypertrophic scarring.

As used herein, “liver fibrosis” includes liver fibrosis due to anycause, including but not limited to virally-induced liver fibrosis suchas that due to hepatitis B or C virus; exposure to alcohol (alcoholicliver disease), certain pharmaceutical compounds including but notlimited to methotrexate, some chemotherapeutic agents, and chronicingestion of arsenicals or vitamin A in megadoses, oxidative stress,cancer radiation therapy or certain industrial chemicals including butnot limited to carbon tetrachloride and dimethylnitrosamine; anddiseases such as primary biliary cirrhosis, primary sclerosingcholangitis, fatty liver, obesity, non-alcoholic steatohepatitis, cysticfibrosis, hemochromatosis, auto-immune hepatitis, and steatohepatitis.Current therapy in liver fibrosis is primarily directed at removing thecausal agent, e.g., removing excess iron (e.g., in the case ofhemochromatosis), decreasing viral load (e.g., in the case of chronicviral hepatitis), or eliminating or decreasing exposure to toxins (e.g.,in the case of alcoholic liver disease). Anti-inflammatory drugs such ascorticosteroids and colchicine are also known for use in treatinginflammation that can lead to liver fibrosis. As is known in the art,liver fibrosis may be clinically classified into five stages of severity(S0, S1, S2, S3, and S4), usually based on histological examination of abiopsy specimen. S0 indicates no fibrosis, whereas S4 indicatescirrhosis. While various criteria for staging the severity of liverfibrosis exist, in general early stages of fibrosis are identified bydiscrete, localized areas of scarring in one portal (zone) of the liver,whereas later stages of fibrosis are identified by bridging fibrosis(scarring that crosses zones of the liver).

In one embodiment, the invention relates to a method of treating orpreventing organ fibrosis in a subject, comprising administering to thesubject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the fibrosis is liver fibrosis.

In one embodiment, the invention relates to a method of treating orpreventing gastrointestinal disease in a subject, comprisingadministering to the subject in need thereof an effective amount of acompound of the invention or a pharmaceutically acceptable salt,solvate, or amino acid conjugate thereof. In one embodiment, theinvention relates to a method of treating gastrointestinal disease. Inone embodiment, the invention relates to a method of preventinggastrointestinal disease. In one embodiment, the gastrointestinaldisease is selected from inflammatory bowel disease (IBD), irritablebowel syndrome (IBS), bacterial overgrowth, malabsorption,post-radiation colitis, and microscopic colitis. In one embodiment, theinflammatory bowel disease is selected from Crohn's disease andulcerative colitis.

In one embodiment, the invention relates to a method of treating orpreventing renal disease in a subject, comprising administering to thesubject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating renal disease. In one embodiment, the invention relates to amethod of preventing renal disease. In one embodiment, the renal diseaseis selected from diabetic nephropathy, focal segmentalglomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronicglomerulonephritis, chronic transplant glomerulopathy, chronicinterstitial nephritis, and polycystic kidney disease.

In one embodiment, the invention relates to a method of treating orpreventing metabolic disease in a subject, comprising administering tothe subject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating renal disease. In one embodiment, the invention relates to amethod of preventing renal disease. In one embodiment, the metabolicdisease is selected from insulin resistance, hyperglycemia, diabetesmellitus, diabesity, and obesity. In one embodiment, the diabetesmellitus is type I diabetes. In one embodiment, the diabetes mellitus istype II diabetes.

Diabetes mellitus, commonly called diabetes, refers to a disease orcondition that is generally characterized by metabolic defects inproduction and utilization of glucose which result in the failure tomaintain appropriate blood sugar levels in the body.

In the case of type II diabetes, the disease is characterized by insulinresistance, in which insulin loses its ability to exert its biologicaleffects across a broad range of concentrations. This resistance toinsulin responsiveness results in insufficient insulin activation ofglucose uptake, oxidation and storage in muscle and inadequate insulinrepression of lipolysis in adipose tissue and of glucose production andsecretion in liver. The resulting condition is elevated blood glucose,which is called “hyperglycemia”. Uncontrolled hyperglycemia isassociated with increased and premature mortality due to an increasedrisk for microvascular and macrovascular diseases, including retinopathy(the impairment or loss of vision due to blood vessel damage in theeyes); neuropathy (nerve damage and foot problems due to blood vesseldamage to the nervous system); and nephropathy (kidney disease due toblood vessel damage in the kidneys), hypertension, cerebrovasculardisease, and coronary heart disease. Therefore, control of glucosehomeostasis is a critically important approach for the treatment ofdiabetes.

Insulin resistance has been hypothesized to unify the clustering ofhypertension, glucose intolerance, hyperinsulinemia, increased levels oftriglyceride and decreased HDL cholesterol, and central and overallobesity. The association of insulin resistance with glucose intolerance,an increase in plasma triglyceride and a decrease in high-densitylipoprotein cholesterol concentrations, hypertension, hyperuricemia,smaller denser low-density lipoprotein particles, and higher circulatinglevels of plasminogen activator inhibitor-1, has been referred to as“Syndrome X”. Accordingly, methods of treating or preventing anydisorders related to insulin resistance including the cluster of diseasestates, conditions or disorders that make up “Syndrome X” are provided.In one embodiment, the invention relates to a method of treating orpreventing metabolic syndrome in a subject, comprising administering tothe subject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating metabolic syndrome. In another embodiment, the inventionrelates to a method of preventing metabolic syndrome.

In one embodiment, the invention relates to a method of treating orpreventing cancer in a subject, comprising administering to the subjectin need thereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof. In one embodiment, the invention relates to a method oftreating cancer. In one embodiment, the invention relates to a method ofpreventing cancer. In one embodiment, the cancer is selected fromhepatocellular carcinoma, colorectal cancer, gastric cancer, renalcancer, prostate cancer, adrenal cancer, pancreatic cancer, breastcancer, bladder cancer, salivary gland cancer, ovarian cancer, uterinebody cancer, and lung cancer. In one embodiment, the cancer ishepatocellular carcinoma. In one embodiment, the cancer is colorectalcancer. In one embodiment, the cancer is gastric cancer. In oneembodiment, the cancer is renal cancer. In one embodiment, the cancer isprostate cancer. In one embodiment, the cancer is adrenal cancer. In oneembodiment, the cancer is pancreatic cancer. In one embodiment, thecancer is breast cancer. In one embodiment, the cancer is bladdercancer. In one embodiment, the cancer is salivary gland cancer. In oneembodiment, the cancer is ovarian cancer. In one embodiment, the canceris uterine body cancer. In one embodiment, the cancer is lung cancer.

In another embodiment, at least one of an agent selected from Sorafenib,Sunitinib, Erlotinib, or Imatinib is co-administered with the compoundof the invention to treat cancer. In one embodiment, at least one of anagent selected from abarelix, aldeleukin, allopurinol, altretamine,amifostine, anastozole, bevacizumab, capecitabine, carboplatin,cisplatin, docetaxel, doxorubicin, erlotinib, exemestane,5-fluorouracil, fulvestrant, gemcitabine, goserelin acetate, irinotecan,lapatinib ditosylate, letozole, leucovorin, levamisole, oxaliplatin,paclitaxel, panitumumab, pemetrexed disodium, profimer sodium,tamoxifen, topotecan, and trastuzumab is co-administered with thecompound of the invention to treat cancer.

Appropriate treatment for cancers depends on the type of cell from whichthe tumor derived, the stage and severity of the malignancy, and thegenetic abnormality that contributes to the tumor.

Cancer staging systems describe the extent of cancer progression. Ingeneral, the staging systems describe how far the tumor has spread andputs patients with similar prognosis and treatment in the same staginggroup. In general, there are poorer prognoses for tumors that havebecome invasive or metastasized.

In one type of staging system, cases are grouped into four stages,denoted by Roman numerals I to IV. In stage I, cancers are oftenlocalized and are usually curable. Stage II and IIIA cancers are usuallymore advanced and may have invaded the surrounding tissues and spread tolymph nodes. Stage IV cancers include metastatic cancers that havespread to sites outside of lymph nodes.

Another staging system is TNM staging which stands for the categories:Tumor, Nodes, and Metastases. In this system, malignancies are describedaccording to the severity of the individual categories. For example, Tclassifies the extent of a primary tumor from 0 to 4 with 0 representinga malignancy that does not have invasive activity and 4 representing amalignancy that has invaded other organs by extension from the originalsite. N classifies the extent of lymph node involvement with 0representing a malignancy with no lymph node involvement and 4representing a malignancy with extensive lymph node involvement. Mclassifies the extent of metastasis from 0 to 1 with 0 representing amalignancy with no metastases and 1 representing a malignancy withmetastases.

These staging systems or variations of these staging systems or othersuitable staging systems may be used to describe a tumor such ashepatocellular carcinoma. Few options only are available for thetreatment of hepatocellular cancer depending on the stage and featuresof the cancer. Treatments include surgery, treatment with Sorafenib, andtargeted therapies. In general, surgery is the first line of treatmentfor early stage localized hepatocellular cancer. Additional systemictreatments may be used to treat invasive and metastatic tumors.

In one embodiment, the invention relates to a method of treating orpreventing gallstones in a subject, comprising administering to thesubject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating gallstones. In one embodiment, the invention relates to amethod of preventing gallstones.

A gallstone is a crystalline concretion formed within the gallbladder byaccretion of bile components. These calculi are formed in thegallbladder but may distally pass into other parts of the biliary tractsuch as the cystic duct, common bile duct, pancreatic duct, or theampulla of Vater. Rarely, in cases of severe inflammation, gallstonesmay erode through the gallbladder into adherent bowel potentiallycausing an obstruction termed gallstone ileus. Presence of gallstones inthe gallbladder may lead to acute cholecystitis, an inflammatorycondition characterized by retention of bile in the gallbladder andoften secondary infection by intestinal microorganisms, predominantlyEscherichia coli, and Bacteroides species.

Presence of gallstones in other parts of the biliary tract can causeobstruction of the bile ducts, which can lead to serious conditions suchas ascending cholangitis or pancreatitis. In one embodiment, theinvention relates to a method of treating or preventing cholesterolgallstone disease in a subject, comprising administering to the subjectin need thereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, or amino acid conjugatethereof. In one embodiment, the invention relates to a method oftreating cholesterol gallstone disease. In one embodiment, the inventionrelates to a method of preventing cholesterol gallstone disease.

In one embodiment, the invention relates to a method of treating orpreventing neurological disease in a subject, comprising administeringto the subject in need thereof an effective amount of a compound of theinvention or a pharmaceutically acceptable salt, solvate, or amino acidconjugate thereof. In one embodiment, the invention relates to a methodof treating neurological disease. In one embodiment, the inventionrelates to a method of preventing neurological disease. In oneembodiment, the neurological disease is stroke.

In one embodiment, the invention relates to a method as described hereinand further wherein, the compound is administered by a route selectedfrom oral, parenteral, intramuscular, intranasal, sublingual,intratracheal, inhalation, ocular, vaginal, rectal, andintracerebroventricular. In one embodiment, the route is oral.

In one embodiment, the compound utilized in one or more of the methodsdescribed herein is an FXR agonist. In one embodiment, the compound is aselective FXR agonist. In another embodiment, the compound does notactivate TGR5. In one embodiment, the compound does not activate othernuclear receptors involved in metabolic pathways (e.g., as measured byan AlphaScreen assay). In one embodiment, such other nuclear receptorsinvolved in metabolic pathways are selected from LXRβ, PXR, CAR, PPARα,PPARδ, PPARγ, RAR, RARα, VDR, TR, PR, RXR, GR, and ER. In oneembodiment, the compound induces apoptosis.

In one embodiment, the invention relates to a method of regulating theexpression level of one or more genes involved in bile acid homeostasis.

In one embodiment, the invention relates to a method of down regulatingthe expression level of one or more genes selected from CYP7α1 andSREBP-IC in a cell by administering to the cell a compound of theinvention. In one embodiment, the invention relates to a method of upregulating the expression level of one or more genes selected from OSTα,OSTβ, BSEP, SHP, UGT2B4, MRP2, FGF-19, PPARγ, PLTP, APOCII, and PEPCK ina cell by administering to the cell a compound of the invention.

The invention also relates to the manufacture of a medicament fortreating or preventing a disease or condition (e.g., a disease orcondition mediated by FXR), wherein the medicament comprises a compoundof the invention or a pharmaceutically acceptable salt, solvate, oramino acid conjugate thereof. In one embodiment, the invention relatesto the manufacture of a medicament for treating or preventing any one ofthe diseases or conditions described herein above, wherein themedicament comprises a compound of the invention or a pharmaceuticallyacceptable salt, solvate, or amino acid conjugate thereof.

The invention also relates to a composition for use in a method fortreating or preventing a disease or condition (e.g., a disease orcondition mediated by FXR), wherein the composition comprises a compoundof the invention or a pharmaceutically acceptable salt, solvate, oramino acid conjugate thereof. In one embodiment, the invention relatesto a composition for use in a method for treating or preventing any oneof the diseases or conditions described herein above, wherein thecomposition comprises a compound of the invention or a pharmaceuticallyacceptable salt, solvate, or amino acid conjugate thereof.

The methods of the invention comprise the step of administering aneffective amount of a compound of the invention. As used herein, theterm an “effective amount” refers to an amount of a compound of theinvention which is sufficient to achieve the stated effect. Accordingly,an effective amount of a compound of the invention used in a method forthe prevention or treatment of FXR mediated diseases or conditions willbe an amount sufficient to prevent or treat the FXR mediated disease orcondition.

Similarly, an effective amount of a compound of the invention for use ina method for the prevention or treatment of a cholestatic liver diseaseor increasing bile flow will be an amount sufficient to increase bileflow to the intestine.

The amount of the compound of the invention which is required to achievethe desired biological effect will depend on a number of factors such asthe use for which it is intended, the means of administration, and therecipient, and will be ultimately at the discretion of the attendantphysician or veterinarian. In general, a typical daily dose for thetreatment of a FXR mediated disease and condition, for instance, may beexpected to lie in the range of from about 0.01 mg/kg to about 100mg/kg. This dose may be administered as a single unit dose or as severalseparate unit doses or as a continuous infusion. Similar dosages wouldbe applicable for the treatment of other diseases, conditions andtherapies including the prevention and treatment of cholestatic liverdiseases.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id or a pharmaceutically acceptable salt, solvate, oramino acid conjugate thereof, and wherein the disease or condition ismediated by FXR.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the disease is selected from cardiovasculardisease, chronic liver disease, lipid disorder, gastrointestinaldisease, renal disease, metabolic disease, cancer, and neurologicaldisease.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the disease is cardiovascular diseaseselected from atherosclerosis, arteriosclerosis, dyslipidemia,hypercholesteremia, hyperlipidemia, hyperlipoproteinemia, andhypertriglyceridemia.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the disease is chronic liver diseaseselected from primary biliary cirrhosis (PBC), cerebrotendinousxanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug inducedcholestasis, intrahepatic cholestasis of pregnancy, parenteral nutritionassociated cholestasis (PNAC), bacterial overgrowth or sepsis associatedcholestasis, autoimmune hepatitis, chronic viral hepatitis, alcoholicliver disease, nonalcoholic fatty liver disease (NAFLD), nonalcoholicsteatohepatitis (NASH), liver transplant associated graft versus hostdisease, living donor transplant liver regeneration, congenital hepaticfibrosis, choledocholithiasis, granulomatous liver disease, intra- orextrahepatic malignancy, Sjogren's syndrome, Sarcoidosis, Wilson'sdisease, Gaucher's disease, hemochromatosis, and alpha 1-antitrypsindeficiency.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the disease is gastrointestinal diseaseselected from inflammatory bowel disease (IBD), irritable bowel syndrome(IBS), bacterial overgrowth, malabsorption, post-radiation colitis, andmicroscopic colitis.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the inflammatory bowel disease is Crohn'sdisease or ulcerative colitis.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the disease is renal disease selected fromdiabetic nephropathy, focal segmental glomerulosclerosis (FSGS),hypertensive nephrosclerosis, chronic glomerulonephritis, chronictransplant glomerulopathy, chronic interstitial nephritis, andpolycystic kidney disease.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the disease is metabolic disease selectedfrom insulin resistance, hyperglycemia, diabetes mellitus, diabesity,and obesity.

In one of the embodiments, the present disclosure proves a method oftreating or preventing a disease or condition in a subject in needthereof comprising administering an effective amount of the compound offormula I or Ia-Id, wherein the disease is cancer selected fromhepatocellular carcinoma, colorectal cancer, gastric cancer, renalcancer, prostate cancer, adrenal cancer, pancreatic cancer, breastcancer, bladder cancer, salivary gland cancer, ovarian cancer, uterinebody cancer, and lung cancer.

Synthesis of the Compounds of the Invention

The following Schemes and Examples are illustrative and should not beinterpreted in any way so as to limit the scope of the invention.

The present invention provides a method of synthesizing compounds ofFormula I,

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein R¹-R¹⁰, m, n, p, and - - - are as described herein.

The synthetic processes of the invention can tolerate a wide variety offunctional groups, therefore various substituted starting materials canbe used. The processes generally provide the desired final compound ator near the end of the overall process, although it may be desirable incertain instances to further convert the compound to a pharmaceuticallyacceptable salt, solvate or amino acid conjugate thereof.

The compounds of the invention can be prepared in a variety of waysusing commercially available starting materials, compounds known in theliterature, or from readily prepared intermediates, by employingstandard synthetic methods and procedures either known to those skilledin the art, or which will be apparent to the skilled artisan in light ofthe teachings herein. Standard synthetic methods and procedures for thepreparation of organic molecules and functional group transformationsand manipulations can be obtained from the relevant scientificliterature or from standard textbooks in the field. Although not limitedto any one or several sources, classic texts such as Smith, M. B.,March, J., March's Advanced Organic Chemistry: Reactions, Mechanisms,and Structure, 5th edition, John Wiley & Sons: New York, 2001; andGreene, T. W., Wuts, P. G. M., Protective Groups in Organic Synthesis,3rd edition, John Wiley & Sons: New York, 1999, incorporated byreference herein, are useful and recognized reference textbooks oforganic synthesis known to those in the art. The following descriptionsof synthetic methods are designed to illustrate, but not to limit,general procedures for the preparation of compounds of the presentinvention.

All the abbreviations used in this application are found in “ProtectiveGroups in Organic Synthesis” by John Wiley & Sons, Inc., or the MERCKINDEX by MERCK & Co.,

Inc., or other chemistry books or chemicals catalogues by chemicalsvendor such as Aldrich, or according to usage know in the art.

The synthetic process to afford compounds of the invention can be usedaccording to the procedures set forth below in Schemes 1-6.

Pharmacology of the Compounds of the Invention

In general, the potential of a compound of the invention as a drugcandidate may be tested using various assays known in the art. Forexample, for the in-vitro validation of FXR, its activity andselectivity can be evaluated using AlphaScreen (biochemical assay); geneexpression can be evaluated using RT-PCR (FXR target gene); andcytotoxicity (e.g., HepG2) can be evaluated using ATP content LDHrelease, and Caspase-3 activation. For the in-vitro validation for TGR5,its activity and selectivity can be evaluated using HTR-FRET (cell-basedassay); gene expression can be evaluated using RT-PCR (TGR5 target gene(i.e., cFOS)); and cytotoxicity (e.g., HepG2) can be evaluated using ATPcontent, LDH release, and Caspase-3 activation. The following compoundscan be used as controls in the examples below.

As used herein Compound A is

which is also known as obeticholic acid, INT-747, 6-ECDCA, 6-alpha-ethylchenodeoxycholic acid, or 6α-ethyl-3α,7α-dihydroxy-5β-cholan-24-oicacid.

As used herein Compound B is

which is also known as INT-767 or6α-ethyl-3α,7α,23-trihydroxy-24-nor-5β-cholan-23-sulphate sodium salt.

As used herein, Compound C is

which is also known as INT-777 or 6α-ethyl-23(S)-methyl-3α,7α,12αtrihydroxy-5β-cholan-24-oic acid.

As used herein, Compound D

which is also known as 6α-ethyl-23(R)-methyl chenodeoxycholic acid, andS-EMCDCA.

As used herein, Compound E is

As used herein, cholic acid is

which is also known as CA.

As used herein, chenodeoxycholic acid is

which is also known as CDCA.

As used herein, ursodeoxycholic acid is

which is also known as UDCA.

As used herein taurochenodeoxycholic acid is

which is also known as TCDCA.

As used herein, tauroursodeoxycholic acid is

which is also known as TUDCA.

As used herein, lithocholic acid is

which is also known as LCA.

EXAMPLES Example 1 Synthesis of3α,7α,11β-trihydroxy-6α-ethyl-24-nor-5β-cholan-23-oic acid (Compound 1)

Compound 1 was prepared according to the procedures described in Scheme1 and from 6-ethyl-cholic acid (6-ECA, Compound A1) as the startingmaterial. Compound A1 was prepared by methods known in the art. Forexample, Compound A1 can be prepared by the procedures described inPellicciari, R., et al., J. Med. Chem. 2009, 52, 7958-7961.

3α,7α,12α-Trihydroxy-6α-ethyl-5β-bisnorcholanyldiphenylethylene(Compound B1)

A solution of Compound A1 (8 g, 18.32 mmol) and para-toluenesulphonicacid (p-TSA) (352 mg, 1.83 mmol) in MeOH (200 mL) was treated underultrasound for 3 h. The mixture was concentrated under vacuum, dilutedwith CHCl₃, and washed with a saturated solution of NaHCO₃. The organicphase was washed with brine, dried over anhydrous Na₂SO₄, andconcentrated under reduced pressure to obtain 7.96 g of methyl6α-ethylcholate derivative. The methyl ester thus formed (17.66 mmol)was dissolved in freshly distilled THF (80 mL) and the mixture waswarmed up to 50° C. under magnetic stirring and argon atmosphere. PhMgBrin Et₂O (176.6 mmol) was then added dropwise and the resulting mixturewas refluxed for 14 h. The suspension was treated with aqueous HCl (50mL) and extracted with EtOAc (3×120 mL). The collected organic layerswere washed with a saturated solution of NaHCO₃, brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The resultingoil was treated with 160 mL of HCl:EtOH (3:1, v/v), refluxed for 3 h andstirred at room temperature overnight. EtOH was removed under vacuum andthe mixture was extracted with EtOAc (3×100 mL). The combined organicphases were washed with a saturated solution of NaHCO₃, brine, driedover anhydrous Na₂SO₄, and concentrated under reduced pressure. Thecrude was purified by silica gel flash chromatography to obtain thedesired product B1 in 76% yield (7.7 g, 13.85 mmol).

3α,7α-Diacetoxy-12-oxo-6α-ethyl-5β-bisnorcholanyldiphenylethylene(Compound C1)

A solution of Compound B1 (7.7 g, 13.85 mmol) and p-TSA (266 mg, 1.38mmol) in MeOAc (70 mL) was refluxed for 2 d. The mixture was washed witha saturated solution of NaHCO₃, brine, dried over anhydrous Na₂SO₄, andconcentrated under vacuum to give 8.15 g of3α-acetoxy-7α,12α-dihydroxy-6α-ethyl-5β-bisnorcholanyldiphenylethylene.The crude (8.18 g) was dissolved in dry CH₂Cl₂ (270 mL). Pyridiniumchlorochromate (PCC) (2.95 g) was added and the mixture was stirred for4 h. The resulting brown suspension was filtered, treated with aqueousHCl, and the organic layer was washed with H₂O and brine. After beingdried over anhydrous Na₂SO₄ and concentrated under reduced pressure, thecrude was purified by silica gel flash chromatography to obtain 5.6 g(9.39 mmol) of the desired 12-oxo derivative. The intermediate was thendissolved in CH₂Cl₂ (80 mL), treated with Ac2O (4.5 mL, 46.95 mmol),Bi(OTf)₃ (306 mg, 0.469 mmol), and stirred for 40 min. The suspensionthus obtained was filtered and acidified with aqueous HCl. The organicphase was washed with H₂O and brine, dried over anhydrous Na₂SO₄ andconcentrated under vacuum. The crude was filtered on silica gel pad toobtain 5.3 g (8.29 mmol) of Compound C1 in 60% yield.

Methyl 3αa,7α-diacetoxy-12-oxo-6α-ethyl-24-nor-5β-cholan-23-oate(Compound D1)

NaIO₄ (15.97 g, 74.66 mmol) was stirred in 15 mL of H₂O and 2 N H₂SO₄(2.4 mL). After 1 h the solution was cooled at 0° C., RuCl₃ (85.9 mg,0.415 mmol) was added and the mixture was magnetically stirred for 1 h.MeCN (23.5 mL) was added as phase transfer and after 5 min a solution ofCompound C1 (5.3 g, 8.29 mmol) in EtOAc (32.5 mL) was dropped andallowed to react for 1 h. The mixture was filtered off, poured into H₂O,and extracted with EtOAc (3×100 mL). The combined organic layers werewashed with brine, dried over Na₂SO₄, and concentrated under reducedpressure. The resulting residue was filtered on silica gel pad to give5.33 g of 6α-ethyl-24-nor-cholic acid derivative which was dissolved inMeOH (90 mL), treated under ultrasound in the presence of p-TSA (160 mg,0.829 mmol) for 3 h and then refluxed for 1 h. The mixture wasconcentrated under vacuum, diluted with CHCl₃, and washed with asaturated solution of NaHCO₃. The organic phase was washed with brine,dried over anhydrous Na₂SO₄, and concentrated under vacuum. The crudewas purified by silica gel flash chromatography to give Compound D1 in86% yield (3.73 g, 7.19 mmol).

Methyl11α-bromine-3α,7α-diacetoxy-12-oxo-6α-ethyl-24-nor-5β-cholan-23-oate(Compound E1)

A solution of Br₂ in anhydrous benzene (2 M, 4.67 mL) was added dropwiseto a solution of Compound D1 (3.73 g, 7.19 mmol) in benzene (156 mL).The resulting red solution was allowed to react at 30° C. under argonatmosphere for 3 days. The mixture was poured into aqueous solution ofNa₂S₂O₃ and the yellow suspension extracted with EtOAc (3×100 mL). Thecollected organic layers were washed with H₂O and brine, dried overanhydrous Na₂SO₄, and concentrated under vacuum. The crude was purifiedby silica gel flash chromatography to obtain Compound E1 as white-yellowsolid (2.65 g, 4.43 mmol).

Methyl 3α,7α-diacetoxy-11β,12β-oxo-6α-ethyl-24-nor-5β-cholan-23-oate(Compound F1)

NaOAc (2.65 g, 32.81 mmol) and NaBH₄ (808 mg, 21.27 mmol) were added toa solution of Compound E1 (2.65 g, 4.43 mmol) in freshly distilledpyridine (27.5 mL) and the suspension was allowed to react at 25° C.under Na atmosphere for 14 h. The mixture was treated with aqueous HCland extracted with EtOAc (3×80 mL). The combined organic phases werewashed with H₂O, brine, and dried under vacuum. The crude oil waspurified by silica gel flash chromatography to obtain 1.52 g (2.85 mmol)of Compound F1 in 64% yield.

Methyl12α-iodine-3α,7α-diacetoxy-11-oxo-6α-ethyl-24-nor-5β-cholan-23-oate(Compound G1)

To a solution of Compound F1 (1.52 g, 2.85 mmol) in AcOH (40 mL), HI 57%(3.6 g, 28.5 mmol) was added dropwise and the mixture was allowed toreact at room temperature for 30 min. The mixture was treated with anaqueous solution of NaHSO₃, poured into iced H₂O, filtered, and theresulting solid dissolved in AcOH (35 mL). A solution of CrO₃ (1.4 g,14.3 mmol) in AcOH (40 mL) and H₂O (8 mL) was added dropwise and themixture stirred for 45 min. The reaction was quenched with an aqueoussolution of NaHSO₃ and poured into iced water. The suspension wasfiltered and the solid dissolved in CHCl₃. The solution was then washedwith H₂O, brine, dried over anhydrous Na₂SO₄, and concentrated underreduced pressure. The crude was purified by silica gel flashchromatography to give Compound G1 as pure product (1 g, 1.67 mmol).

3α,7α-Dihydroxy-11-oxo-6α-ethyl-24-nor-5β-cholan-23-oic acid (CompoundH₁)

NaOAc (3.8 g, 46.76 mmol) and Zn dust (3.8 g, 58.45 mmol) were added toa solution of Compound G1 (1 g, 1.69 mmol) in AcOH (30 mL) and theresulting suspension was refluxed for 2 h. The mixture was filtered andthe filtrate treated with H₂O at 0° C. up to precipitation. Theprecipitate was dissolved in CHCl₃ and the aqueous phase extracted withCHCl₃ (3×50 mL). The collected organic layers were treated with asaturated solution of NaHCO₃, washed with brine, dried over anhydrousNa₂SO₄ and then concentrated under vacuum. The crude (880 mg) wasdissolved in MeOH and H₂O, NaOH (25.45 mmol) was added and the mixturewas refluxed for 36 h. The resulting solution was concentrated underreduced pressure, diluted with H₂O and treated with aqueous HCl. It wasextracted with CHCl₃ (3×50 mL) and the combined organic phases werewashed with brine, dried over anhydrous Na₂SO₄, and concentrated undervacuum to give Compound H1.

3α,7α,11β-Trihydroxy-6α-ethyl-24-nor-5β-cholan-23-oic acid (Compound 1)

To a solution of Compound H1 (650 mg, 1.54 mmol) in THF:H₂O (33 mL, 4:1v/v), NaBH₄ (407 mg, 10.78 mmol) was added portionwise at 0° C. and theresulting suspension was allowed to react at room temperature for 5 h.After being treated with H₂O and aqueous HCl, the crude reaction mixturewas extracted with CHCl₃ (3×50 mL). The collected organic layers werewashed with brine, dried over anhydrous Na₂SO₄, and concentrated undervacuum to give 650 mg of Compound 1 (1.69 mmol, quantitative yield) (9%overall yield from 1).

Compound 1: ¹H-NMR (400 MHz, CD3OD): δ 0.89 (3H, t, J=7.33 Hz, CH₃-25),0.94 (3H, s, CH₃-18), 1.04 (3H, d, J=5.46 Hz, CH₃-21), 1.13 (3H, s,CH₃-19), 3.30-3.35 (1H, m, CH-3), 3.71 (1H, s, CH-7), 4.19 (1H, s,CH-11). ¹³C-NMR (400 MHz, CD₃OD): 12.0, 14.6, 19.9, 23.5, 24.6, 27.7,29.1, 31.9, 34.7, 35.2, 36.4, 36.9, 38.3 (2×), 42.6 (2 x), 42.8, 49.5,49.9, 52.2, 57.9, 69.0, 71.4, 73.3, 177.7.

Example 2 Synthesis of3α,7α,11β-trihydroxy-6α-ethyl-24-nor-5β-cholan-23-ol (Compound 2)

Compound 2 was prepared according to the procedures set forth in Scheme2. Compound 2 was prepared from Compound 1 as the starting material.

Example 3 Synthesis of3α,7α,11β,23-tetrahydroxy-6α-ethyl-24-nor-5β-cholan-23-O-sulphate sodiumsalt (Compound 3)

Compound 3 was prepared according to the procedures set forth in Scheme2. Compound 3 was prepared from Compound 1 as the starting material.

3α-Acetoxy-7α,11β-dihydroxy-6α-ethyl-24-nor-5β-cholan-23-oic acid(Compound J1)

Ac₂O (2.08 mL, 21.6 mmol) was added to a solution of Compound 1 (460 mg,1.08 mmol) in THF (35 mL) and the mixture was refluxed for 18 h. Theresulting solution was treated with aqueous HCl extracted with EtOAc(3×30 mL). The combined organic phases were washed with H₂O, brine,dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Thecrude was purified by silica gel flash chromatography to obtain CompoundJ1 (255 mg, 0.548 mmol).

3α-Acetoxy-7α,11β-dihydroxy-6α-ethyl-24-nor-5β-cholan-23-ol (CompoundK1)

A solution of Compound J1 (250 mg, 0.538 mmol), EtCOCl (0.51 mL, 5.326mmol), and Et₃N (0.81 mL, 5.649 mmol) in THF (7.5 mL) was allowed toreact for 14 h at room temperature. The reaction mixture was thenfiltered, treated with a suspension of NaBH₄ (306 mg, 8.07 mmol) in H₂O(2.5 mL) and stirred for 2 h. The mixture was acidified with aqueous HCland extracted with EtOAc (3×30 mL). The combined organic extracts werewashed with brine, dried over Na₂SO₄, and concentrated under vacuum. Thecrude was filtered on silica gel pad to give Compound K1 (150 mg, 0.333mmol).

3α,7α,11β-trihydroxy-6α-ethyl-24-nor-5β-cholan-23-ol (2) and3α,7α,11β,23-tetrahydroxy-6α-ethyl-24-nor-5β-cholan-23-O-sulphate sodiumsalt (Compound 3)

To a solution of Compound K1 (150 mg, 0.333 mmol) in pyridine (6 mL),was added PyrSO₃ (133 mg, 0.832 mmol) and the resulting mixture wasstirred under argon atmosphere for 24 h. The reaction mixture wasdiluted with H₂O (2 mL) and concentrated at reduced pressure to removepyridine. The residue was treated with a solution of NaOH (200 mg, 4.995mmol) in MeOH:H₂O (10 mL) and refluxed overnight. The mixture was driedunder vacuum to remove MeOH, diluted with H₂O (2 mL), and washed withEt₂O (3×20 mL): the combined ethereal phases were washed with brine,dried over anhydrous Na₂SO₄, and purified by flash chromatography togive the 3α,7α,11β-trihydroxy-6α-ethyl-24-nor-5β-cholan-23-ol (2) aspure white solid (55 mg, 0.134 mmol). The aqueous alkaline phase wasfiltered on a reverse phase RP-18 pad to obtain Compound 3 as pure whitesolid (60 mg, 0.117 mmol).

Compound 2: ¹H-NMR (400 MHz, CDCl₃): δ 0.88-0.92 (6H, m, CH₃-25,CH₃-18), 0.97 (3H, d, J=6.5 Hz, CH₃-21), 1.14 (3H, s, CH₃-19), 3.40-3.47(1H, m, CH-3), 3.62-3.72 (2H, m, CH₂-23), 3.80 (1H, s, CH-7), 4.25 (1H,d, J=2.72 Hz, CH-11). ¹³C-NMR (400 MHz, CDCl₃): 11.6, 14.4, 18.8, 22.2,23.8, 27.0, 28.0, 31.1, 32.9, 34.1, 35.3, 35.7, 36.4, 37.1, 38.8, 40.6,41.6, 47.7, 48.8, 50.9, 56.8, 60.7, 68.8, 71.0, 72.3.

Compound 3: ¹H-NMR (400 MHz, CD₃OD): δ 0.90-0.94 (6H, m, CH₃-25,CH₃-18), 1.04 (3H, d, J=6.4 Hz, CH₃-21), 1.15 (3H, s, CH₃-19), 3.32-3.40(1H, m, CH-3), 3.74 (1H, s, CH-7), 4.02-4.08 (2H, m, CH₂-23), 4.21 (1H,s, CH-11). ¹³C-NMR (400 MHz, CD₃OD): 12.0, 14.6, 19.1, 23.5, 24.7, 27.7,29.1, 31.9, 34.3, 34.8, 36.4, 36.5, 36.9, 38.3 (×2), 42.6, 42.8, 49.5,50.0, 52.2, 58.2, 67.2, 69.0, 71.4, 73.3.

Example 4 Synthesis of 3α,7α,11β-trihydroxy-6α-ethyl-5β-cholan-24-ol(Compound 4)

4 was prepared according to the procedures set forth in Scheme 3. Thesynthesis of 4 was prepared from Compound L1 as the starting material.Compound L1 was prepared by methods known in the art. For example,Compound L1 can be prepared by the procedures described in U.S.Publication No. 2014/0371190.

3α,7α,11β-Trihydroxy-6αa-ethyl-5β-cholan-24-ol (Compound 4)

A solution of Compound L1 (25 mg, 0.057 mmol) in THF (2 mL) was addeddropwise to a suspension of LiAlH₄ (21.8 mg, 0.572 mmol) in THF (1 mL)cooled at 0° C. The resulting mixture was allowed to react under argonatmosphere and room temperature for 12 h. The suspension was dilutedwith EtOAc (5 mL), treated firstly with H₂O, then with aqueous HCl, andfinally extracted with EtOAc (3×5 mL). The combined organic layers werewashed with brine, dried over anhydrous Na₂SO₄, and concentrated undervacuum. The crude was purified by flash chromatography to give Compound4 as pure white solid (21 mg, 0.051 mmol, 90% yield).

Compound 4: ¹H-NMR (400 MHz, CD₃OD): δ 0.88-0.92 (6H, m, CH₃-26,CH₃-18), 1.00 (3H, d, J=6.25 Hz, CH₃-21), 1.14 (3H, s, CH₃-19),3.31-3.40 (1H, m, CH-3), 3.48-3.55 (2H, m, CH₂-24), 3.73 (1H, s, CH-7),4.19 (1H, s, CH-11). ¹³C-NMR (400 MHz, CD₃OD): 12.0, 14.6, 19.2, 23.5,24.7, 27.7, 29.1, 30.3, 31.9, 33.2, 34.8, 36.4, 36.9, 37.2, 38.3 (×2),42.6, 42.7, 49.5, 50.1, 52.2, 58.1, 63.6, 69.1, 71.4, 73.3.

Example 5 Synthesis of3α,7α,11β-rihydroxy-6α-ethyl-5β-cholan-24-O-sulphate, sodium salt(Compound 5)

Compound 5 was prepared according to the procedures set forth in Scheme4. The synthesis of Compound 5 was prepared from Compound M1 as thestarting material. Compound M1 was prepared by methods known in the art.For example, Compound M1 can be prepared by the procedures described inU.S. Publication No. 2014/0371190.

3α-Acetoxy-7α-hydroxy-11-oxo-6α-ethyl-5β-cholan-24-ol (Compound N1)

To a solution of Compound M1 (120 mg, 0.27 mmol) in freshly distilledTHF (4 mL), NaHCO₃ (417 mg, 4.97 mmol) and Ac₂O (0.47 mL, 4.97 mmol)were added and the suspension was refluxed for 24 h under argonatmosphere. The mixture was cooled to room temperature, treated withaqueous HCl and extracted with EtOAc (3×10 mL). The collected organicphases were sequentially washed with aqueous HCl, water, a saturatedsolution of NaHCO₃, brine, and dried over anhydrous Na₂SO₄. After beingconcentrated under reduced pressure, the crude was dissolved in freshlydistilled THF (3 mL), treated with Et₃N (0.22 mL, 1.54 mmol) and ClCO₂Et(0.14 mL, 1.45 mmol) and the mixture was allowed to react at roomtemperature for 2 h under argon atmosphere. The suspension was filteredand the filtrate treated with a suspension of NaBH₄ (125 mg, 3.30 mmol)in H₂O (1 mL) and stirred for 3 h. The mixture was acidified withaqueous HCl and extracted with EtOAc (3×10 mL). The combined organiclayers were washed with H₂O, brine, dried over anhydrous Na₂SO₄, andconcentrated under vacuum. The crude was purified by flashchromatography thus obtaining Compound N1 (70 mg, 0.15 mmol).

3α-Acetoxy-7α,11β-dihydroxy-6α-ethyl-5β-cholan-24-ol (Compound P1)

To a solution of Compound N1 (0.15 mmol) in a binary mixture of THF andH₂O, NaBH₄ (3.75 mmol) was added and the mixture was stirred at roomtemperature for 24 h. The suspension was treated with aqueous HCl andextracted with EtOAc (3×10 mL). The collected organic phases were washedwith H₂O, brine, dried over anhydrous Na₂SO₄, and concentrated underreduced pressure giving Compound P1 in quantitative yield.

3α,7α,11β-Trihydroxy-6α-ethyl-5β-cholan-24-O-sulphate, sodium salt(Compound 5)

PyrSO₃ (48 mg, 0.30 mmol) was added to a solution of Compound P1 (70 mg,0.15 mmol) in pyridine (2.7 mL) and allowed to react at room temperaturefor 30 h under argon atmosphere. Pyridine was removed under vacuum andthe residue stirred with a solution of NaOH (60 mg, 1.5 mmol) in amixture of MeOH and H₂O for 3 d. The mixture was concentrated underreduced pressure to evaporate MeOH, diluted with H₂O (2 mL), and washedwith Et₂O (3×10 mL). The aqueous alkaline phase was filtered on areverse phase RP-18 pad to obtain Compound 5 (47 mg, 0.085 mmol, 57%yield) as pure white solid.

Compound 5: ¹H-NMR (400 MHz, CD₃OD): δ 0.88-0.92 (6H, m, CH₃-18,CH₃-26), 1.00 (3H, d, J=6.3 Hz, CH₃-21), 1.14 (3H, s, CH₃-19), 3.32-3.35(1H, brm, CH-3), 3.72 (1H, brs, CH-7), 3.94-3.97 (2H, brm, CH₂-24), 4.20(1H, brs, CH-11). ¹³C-NMR (400 MHz, CD₃OD): 12.1, 14.7, 19.1, 23.6,24.7, 27.1, 27.7, 29.1, 31.9, 33.1, 34.8, 36.4, 36.9, 37.0, 38.3 (×2),42.6, 42.7, 49.5, 50.1, 52.2, 58.0, 69.1, 69.7, 71.4, 73.3.

Example 6 Synthesis of3α,7α,11β-trihydroxy-6α-ethyl-22-(1,2,4-oxadiazol-5-oxo-3-yl)-23,24-bisnor-5β-cholane(Compound 6)

3α,7α,11β-Triacetoxy-6α-ethyl-5β-cholan-24-oic acid (Compound Q1)

To a suspension of Compound L1 (660 mg, 1.5 mmol) in CH₂Cl₂ (15 mL),Ac₂O (22.7 mmol) and Bi(OTf)₃ (0.08 mmol) were added and the mixture wasstirred at room temperature for 1 h. The reaction mixture was filteredand the filtrate treated with HCl 37%. The organic phase was washed withH₂O, with a saturated solution of NaHCO₃ and brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to obtain Q1 (820 mg,1.46 mmol, 96% yield), which was used for the next step without furtherpurification.

Compound Q1: ¹H-NMR (400 MHz, CDCl₃): δ 0.76 (3H, s, CH₃-18), 0.87-0.90(6H, m, CH₃-21, CH₃-26), 1.04 (3H, s, CH₃-19), 2.03-2.05 (6H, m, OCOCH₃×2), 2.08 (3H, s, OCOCH₃), 4.52-4.61 (1H, m, CH-3), 5.20 (1H, s, CH-7),5.25 (1H, s, CH-11).

3α,7α,11β-Triacetoxy-6α-ethyl-24-nor-5β-cholan-23-nitrile (Compound R1)

A suspension of Compound Q1 (820 mg, 1.46 mmol) in TFA (4.6 mL) at 0° C.was treated with TFAA (1.55 mL) and stirred at 0° C. for 45 min. NaNO₂(4.4 mmol) was added and the mixture was reacted at 0° C. for 45 min andat 50° C. for additional 45 min. The reaction mixture was cooled to roomtemperature and poured into crushed ice. The aqueous phase was filteredunder vacuum and the resulting orange-yellow solid was dissolved inEtOAc (30 mL), washed with a saturated solution of NaHCO₃, H₂O andbrine. The organic layer was dried over anhydrous Na₂SO₄, concentratedunder reduced pressure and to obtain Compound R1 (770 mg) as a crudethat was used for the next step without further purification.

Compound R1: ¹⁻H-NMR (400 MHz, CDCl₃): δ 0.76 (3H, s, CH₃-18), 0.84-0.87(3H, m, CH₃-25), 1.02 (3H, s, CH₃-19), 1.08 (3H, d, J=6.4 Hz, CH₃-21),2.01 (6H, brs, OCOCH₃ ×2), 2.07 (3H, s, OCOCH₃), 4.52-4.61 (1H, m,CH-3), 5.18 (1H, s, CH-7), 5.24 (1H, s, CH-11).

3α,7α,11β-Trihydroxy-6α-ethyl-24-nor-5β-cholan-23-nitrile (Compound S1)

Compound R1 (770 mg) was dissolved in MeOH (10 mL) and refluxed for 3 dwith NaOH (1.2 g). After solvent removal, the residue was dissolved inCHCl₃ (30 mL) and treated with 1 N HCl. The aqueous phase was extractedwith CHCl₃ and the combined organic layers were washed with H₂O andbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude was purified by silica gel flash chromatographyusing CH₂Cl₂ and MeOH as eluting solvents to obtain Compound S1 (180 mg,0.445 mmol) in high purity grade.

Compound S1: ¹H-NMR (400 MHz, CDCl₃): δ 0.87-0.91 (6H, m, CH₃-18,CH₃-25), 1.13 (3H, s, CH₃-19), 1.17 (3H, d, J=6.5 Hz, CH₃-21), 3.42-3.50(1H, m, CH-3), 3.77 (1H, s, CH-7), 4.28 (1H, s, CH-11).

3α,7α,11β-Trihydroxy-6α-ethyl-24-nor-N-hydroxy-5β-cholan-23-amidine(Compound T1)

NH₂OH.HCl (557 mg) and Na₂CO₃.10H₂O (2.30 g) were added to a solution ofCompound S1 (180 mg, 0.445 mmol) in EtOH (8 mL) and the resultingmixture was refluxed for 2 d. The suspension was cooled to roomtemperature and filtered under vacuum. The solid was washed with EtOAcand the organic phase was washed with H₂O, brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The crude, containingthe desired intermediate Compound T1, was used for the next step withoutfurther purification.

Compound T1: ¹H-NMR (400 MHz, CD₃OD): δ 0.90 (3H, t, J=7.3 Hz, CH₃-25),0.96-0.98 (6H, m, CH₃-18, CH₃-21), 1.14 (3H, s, CH₃-19), 3.31-3.40 (1H,m, CH-3), 3.72 (1H, s, CH-7), 4.20 (1H, s, CH-11).

3α,7α,11β-Trihydroxy-6α-ethyl-24-nor-N-[(ethoxycarbonyl)oxy]-5β-cholan-23-amidine(Compound U1)

To a solution of Compound T1, (180 mg) in THF (2 mL) and pyridine (50μL, 0.6 mmol) cooled at 0° C., a solution of ClCO₂Et (0.45 mmol) in THF(1 mL) was added dropwise and the resulting suspension was stirred underargon atmosphere for 30 min. The mixture was treated with H₂O andextracted with EtOAc (3×10 mL). The collected organic phases were washedwith brine, dried over anhydrous Na₂SO₄, concentrated under vacuum toprovide Compound U1, which was used for the next step without furtherpurification.

Compound U1: ¹H-NMR (400 MHz, CDCl₃): δ 0.86-0.92 (6H, m, CH₃-25,CO₂CH₂CH₃), 1.00 (3H, d, J=6.1 Hz, CH₃-21), 1.11 (3H, s, CH₃-18), 1.23(3H, s, CH₃-19), 3.35-3.44 (1H, m, CH-3), 3.77 (1H, s, CH-7), 4.13-4.39(3H, m, CH-11, CO₂CH₂CH₃), 4.90-5.05 (1H, m, NH).

3α,7α,11β-Trihydroxy-6α-ethyl-22-(1,2,4-oxadiazol-5-oxo-3-yl)-23,24-bisnor-5β-cholane(Compound 6)

Compound U1 (210 mg, 0.412 mmol) was dissolved in toluene (6 mL) andpyridine (0.6 mL) and refluxed under argon atmosphere for 20 h. Afterbeing cooled to room temperature, the mixture was diluted with EtOAc (10mL) and washed with 1 N HCl, H₂O, a saturated solution of NaHCO₃ andbrine. The organic layer was dried over anhydrous Na₂SO₄, concentratedunder reduced pressure and purified to obtain Compound 6 (35 mg,0.076mmol).

Compound 6: ¹H-NMR (400 MHz, CD₃OD): δ 0.89-0.93 (3H, t, J=7.3 Hz,CH₃-24), 0.96-0.99 (6H, m, CH₃-18, CH₃-21), 1.15 (3H, s, CH₃-19), 2.09(1H, d, J=14.1 Hz), 2.37 (1H, d, J=12.3 Hz), 2.57 (1H, d, J=13.4 Hz),3.31-3.40 (1H, m, CH-3), 3.66 (1H, s, OH), 3.73 (1H, s, CH-7), 4.21 (1H,s, CH-11). ¹³C-NMR (100.6 MHz, CD₃OD): δ 12.0, 14.7, 19.2, 23.5, 24.7,27.6, 29.1, 31.9, 34.5, 34.7, 36.1, 36.4, 36.9, 38.2 (×2), 42.6, 43.0,49.4, 49.9, 52.2, 58.4, 68.9, 71.3, 73.3, 169.3, 173.0.

Example 7 Synthesis of3α,7α,11β-trihydroxy-6α-ethyl-23-(1,2,4-oxadiazol-5-oxo-3-yl)-23-nor-5β-cholane(Compound 7)

Methyl 6α-ethyl-3α,7α,11β-trimethoxymethyloxy-5β-cholan-24-oate(Compound V1)

A solution of Compound L1 (730 mg, 1.7 mmol) and p-TSA (0.17 mmol) inMeOH (10 mL) was treated under ultrasounds for 3 h. The solvent wasremoved, the residue dissolved in EtOAc (10 mL) and washed with asaturated solution of NaHCO₃. The aqueous phase was extracted with EtOAc(2×10 mL) and the combined organic layers washed with brine, dried overanhydrous Na₂SO₄ and concentrated under vacuum. The crude (700 mg, 1.55mmol) was dissolved in CH₂Cl₂ (20 mL) and refluxed with DIPEA (18.6mmol), DMAP (0.16 mmol) and MOMCl (15.5 mmol) for 3 d. The mixture wascooled to room temperature and sequentially washed with a saturatedsolution of NH₄C₁, H₂O and brine. The organic phase was dried overanhydrous Na₂SO₄, concentrated under reduced pressure and the obtainedcrude Compound VI which was used for the next step without furtherpurification.

Compound V1: ¹H-NMR (400 MHz, CDCl₃): δ 0.81 (3H, s, CH₃-18), 0.85-0.89(3H, m, CH₃-26), 0.93 (3H, d, J=6.2 Hz, CH₃-21), 1.10 (3H, s, CH₃-19),3.34-3.40 (10H, m, CH-3, OCH₂OCH₃ ×3), 3.53 (1H, s, CH-7), 3.65 (3H, s,CO₂CH₃), 3.93 (1H, s, CH-11), 4.55-4.70 (6H, m, OCH₂OCH₃ ×3).

6α-Ethyl-3α,7α,11β-trimethoxymethyloxy-5β-cholan-24-amide (Compound W1)

To a solution of Compound V1 (980 mg, 1.6 mmol) in MeOH (10 mL), NaOH(15.5 mmol) was added and the mixture was allowed to react at 50° C. Thesolvent was removed under vacuum, the residue dissolved in H₂O (5 mL)and treated with HCl 1 N. The suspension was extracted with CHCl₃ (3×10mL) and the combined organic phases were washed with H₂O and brine,dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Thecrude (840 mg) was dissolved in freshly distilled THF (18 mL), cooled at0° C. and stirred with Et₃N (0.288 mL) and ClCO₂iBu (0.250 mL) for 20min under argon atmosphere. NH₄OH 30% (0.28 mL) was added and theresulting suspension was reacted for 40 min at room temperature. Themixture was treated with H₂O and extracted with EtOAc (3×10 mL). Thecollected organic phases were washed with HCl 1 N, H₂O, a saturatedsolution of NaHCO₃, H₂O and brine, dried over anhydrous Na₂SO₄, andconcentrated under reduced pressure to obtain Compound W1 (900 mg) thatwas used for the next step without further purification.

Compound W1: ¹H-NMR (400 MHz, CDCl₃): δ 0.62-0.82 (9H, m, CH₃-18,CH₃-21, CH₃-26), 0.99 (3H, s, CH₃-19), 3.22-3.30 (10H, m, CH-3, OCH₂OCH₃×3), 3.42 (1H, s, CH-7), 3.82 (1H, s, CH-11), 4.46-4.57 (6H, m, OCH₂OCH₃×3), 6.03 (1H, brs, CONH₂), 6.27 (1H, brs, CONH₂).

6α-Ethyl-3α,7α,11β-trimethoxymethyloxy-5β-cholan-24-nitrile (CompoundX1)

A solution of Compound W1 (890 mg) and CNCl (578 mg) in DMF (22 mL) wasstirred at room temperature under argon atmosphere for 12 h. Theresulting suspension was diluted with EtOAc (50 mL) and washed with H₂O(3×15 mL). The organic layer was washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The crude was purifiedby flash chromatography on silica gel by using PET-EtOAc as elutingsolvent system to obtain Compound X1 as a pale yellow oil (260 mg, 0.473mmol).

Compound X1: ¹H-NMIR (400 MHz, CDCl₃): δ 0.83-0.88 (6H, m, CH₃-18,CH₃-26), 0.95 (3H, d, J=6.0 Hz, CH₃-21), 1.05 (3H, s, CH₃-19), 3.33-3.40(10H, m, CH-3, OCH₂OCH₃ ×3), 3.52 (1H, s, CH-7), 3.91 (1H, s, CH-11),4.55-4.69 (6H, m, OCH₂OCH₃ ×3).

6α-Ethyl-3α,7α,11β-trimethoxymethyloxy-N-hydroxy-5β-cholan-24-amidine(Compound Y1)

NH₂OH.HCl (386 mg) and Na₂CO₃. (1.6 g) were added to a solution ofCompound X1 (170 mg, 0.309 mmol) in EtOH (6 mL) and refluxed tillstarting material consumption. The suspension was cooled to roomtemperature and filtered under vacuum. The remaining solid was washedwith EtOAc (15 mL) and the filtered organic phase was washed with H₂O,brine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude, containing the desired intermediate Compound Y1,was used for the next step without further purification.

Compound Y1: ¹H-NMR (400 MHz, CDCl₃): δ 0.77 (3H, s, CH₃-18), 0.81-0.84(3H, t, J=6.8 Hz, CH₃-26), 0.90 (3H, d, J=6.2 Hz, CH₃-21), 1.05 (3H, s,CH₃-19), 3.30-3.36 (10H, m, CH-3, OCH₂OCH₃ ×3), 3.48 (1H, s, CH-7), 3.88(1H, s, CH-11), 4.51-4.65 (6H, m, OCH₂OCH₃ ×3), 4.76 (2H, brs, NH,NH—OH).

6α-Ethyl-3α,7α,11α-trimethoxymethyloxy-N-[(ethoxycarbonyl)oxy]-5β-cholan-24-amidine(Compound Z1)

To a solution of Compound Y1 (200 mg) in THF (2 mL) and pyridine (0.46mmol) cooled at 0° C., a solution of ClCO₂Et (0.34 mmol) in THF (1 mL)was added dropwise and the resulting suspension was stirred under argonatmosphere for 30 min. The mixture was treated with H₂O and extractedwith EtOAc (3×10 mL). The collected organic phases were washed withbrine, dried over anhydrous Na₂SO₄, concentrated under vacuum to provideCompound Z1, which was used for the next step without furtherpurification.

Compound Z1: ¹H-NMR (400 MHz, CDCl₃): δ 0.77 (3H, s, CH₃-18), 0.81-0.84(3H, t, J=6.8 Hz, CH₃-26), 0.92 (3H, d, J=6.1 Hz, CH₃-21), 1.05 (3H, s,CH₃-19), 1.27 (3H, t, J=7.0 Hz, OCH₂CH₃), 3.30-3.36 (10H, m, CH-3,OCH₂OCH₃ ×3), 3.48 (1H, s, CH-7), 3.88 (1H, s, CH-11), 4.21 (2H, q,J=7.0 Hz, OCH₂CH₃), 4.51-4.65 (6H, m, OCH₂OCH₃ ×3), 4.88 (2H, brm, NH,NH—OH).

3α,7α,11β-Trihydroxy-6α-ethyl-23-(1,2,4-oxadiazol-5-oxo-3-yl)-24-nor-5β-cholane(Compound 7)

Compound Z1 (200 mg) obtained from the previous step was dissolved intoluene (5 mL) and pyridine (0.5 mL), refluxed under argon atmospherefor 8 h, and stirred at room temperature for additional 12 h. Themixture was diluted with EtOAc (10 mL) and sequentially washed with HCl1 N, H₂O, a saturated solution of NaHCO₃ and brine. The organic layerwas dried over anhydrous Na₂SO₄ and concentrated under reduced pressure.The resulting crude was dissolved in acetone (15 mL) and stirred withHCl 3 N (1.5 mL) at 40° C. for 6 h. The mixture was diluted with H₂O andthe organic layer was concentrated under reduced pressure. The aqueousphase was then extracted with EtOAc (3×10 mL) and the combined organiclayers treated with a saturated solution of NaHCO₃, washed with brine,dried over anhydrous Na₂SO₄ and concentrated under vacuum. The crude waspurified by flash chromatography to yield Compound 7 (29.6 mg, 0.062mmol) as a white solid.

Compound 7: ¹H-NMR (400 MHz, CD₃OD): δ 0.89-0.93 (6H, m, CH₃-18,CH₃-25), 1.05 (3H, d, J=6.3 Hz, CH₃-21), 1.15 (3H, s, CH₃-19), 2.07-2.11(1H, m), 2.20-2.24 (1H, m), 2.45-2.50 (1H, m, CH-23), 2.58-2.68 (1H, m,CH-23), 3.32-3.36 (1H, m, CH-3), 3.73 (1H, s, CH-7), 4.20 (1H, s,CH-11). ¹³C-NMR (100.6 MHz, CD₃OD): 12.0, 14.6, 18.7, 23.0, 23.5, 24.7,27.7, 29.1, 30.8, 31.9, 33.1, 34.7, 36.4, 36.8, 36.9, 38.3 (×2), 42.6,42.8, 50.0, 52.2, 57.6, 69.0, 71.4, 73.3, 162.2, 162.8.

Example 6 FXR/TGR5 Activity of the Compounds 1-7

In the nucleus, ligand-bound nuclear receptors (NRs) modulate initiationof transcription by directly interacting with the basal transcriptionalmachinery or by contacting bridging factors called coactivators (Onate,et al., Science, 1995, 270, 1354-1357; Wang, et al., J Biol Chem, 1998,273, 30847-30850; and Zhu, et al., Gene Expr, 1996, 6, 185-195). Theligand-dependent interaction of NRs with their coactivators occursbetween activation function 2 (AF-2), located in the receptorligand-binding domain (LBD) and the nuclear receptor boxes (NR box),located on the coactivators (Nolte, et al., Nature, 1998, 395, 137-143).Several lines of evidence have demonstrated that the LXXLL peptidesequence present in the NR box represents a signature motif thatfacilitates the interaction of different proteins with the AF-2 region(Heery, et al., Nature, 1997, 387, 733-736; and Torchia, et al., Nature,1997, 387, 677-684).

AlphaScreen was used with the aim of identifying novel modulators bytaking advantage of the bimolecular interaction prevailing between FXRand the LXXLL motif present in the NR box of the steroid receptorcoactivator 1 (SRC-1).

Human FXR-LBD-GST was incubated with increasing concentrations of theindicated ligands in the presence of biotinylated LXXLL SRC-1 peptide.The AlphaScreen signal increases when the complex receptor-coactivatoris formed. The compounds of this invention are potent FXR agonists. Dataare provided in Tables 1 and 2.

Bile acids (BAs) modulate not only several nuclear hormone receptors,but are also agonists for the G protein-coupled receptor (GPCR) TGR5(Makishima, et al., Science, 1999, 284, 1362-1365; Parks, et al.,Science, 1999, 284, 1365-1368; Maruyama, et al., Biochem Biophys ResCommun, 2002, 298, 714-719; and Kawamata, et al., J Biol Chem, 2003,278, 9435-9440). Signalling via FXR and TGR5 modulates several metabolicpathways, regulating not only BA synthesis and enterohepaticrecirculation, but also triglyceride, cholesterol, glucose, and energyhomeostasis. To evaluate the capacity of a compound of the invention toactivate TGR5, the compound of the invention and other comparisoncompounds were screened for an increase of intracellular cAMP as aread-out for TGR5 activation. Human enteroendocrine NCI-H₇₁₆ cellsconstitutively expressing TGR5 were exposed to increasing concentrationsof a compound of the invention, and intracellular cAMP levels weremeasured by TR-FRET. Lithocholic acid (LCA) was used as positivecontrol. The compounds of this invention show high selectivity for FXRover TGR5. Data are provided in

TABLE 1 FXR/TGR5 Activity of Compounds 1-5 HTR-FRET AlphaScreen (cAMP)Assay Human TGR5 Human FXR (NCI-H716 cells) Compound Ref CDCA = 15 ± 3μM Ref LCA = 7 ± 3 μM Compound 1 0.68 >100 Compound 2 0.23 93 Compound 30.0075 ± 0.0005 83 ± 7 Compound 4 0.264 ± 0.016 13.7 ± 2.3 Compound 50.015 ± 0.004 78 ± 1 Compound A  0.2 ± 0.018 15 ± 5 Compound B 0.03 0.63Compound C 175 0.9

TABLE 2 FXR/TGR5 Activity of Compounds L1, 3, 5, 6, and 7 AlphaScreenHTR-FRET (cAMP) Human FXR Human TGR5 Compound EC₅₀ (μM) EC₅₀ (μM) L10.15 ± 0.5  No activity Compound 3 0.0075 ± 0.0005 83 ± 7 Compound 50.015 ± 0.004 78 ± 1 Compound 6 0.042 ± 0.002 No activity Compound 70.029 ± 0.005 No activity

TABLE 3 FXR agonist activity across human, mouse, rat, and dog orthologsAlphaScreen AlphaScreen AlphaScreen AlphaScreen hFXR mFXR rFXR dFXRCompound EC₅₀ (μM) EC₅₀ (μM) EC₅₀ (μM) EC₅₀ (μM) L1 0.15 ± 0.5  0.99 ±0.05  1.0 ± 0.03 4 ± 1 Compound 3 0.0075 ± 0.0005 0.25 ± 0.04 0.13 ±0.01 0.9 ± 0.1 Compound 5 0.015 ± 0.004 0.12 ± 0.02 0.14 ± 0.02 0.73 ±0.01 Compound 6 0.042 ± 0.002 0.27 ± 0.02 0.24 ± 0.01 0.8 ± 0.1 Compound7 0.029 ± 0.005 0.21 ± 0.01 0.2 ± 0.01  0.7 ± 0.01

TABLE 4 Cross species TGR5 activity hTGR5 mTGR5 rTGR5 dTGR5 CHO CHO CHOCHO Compound EC₅₀ (μM) EC₅₀ (μM) EC₅₀ (μM) EC₅₀ (μM) L1 No activity Noactivity No activity No activity Compound 3 5 ± 1   3 ± 0.5 No activityNo activity Compound 5 3 ± 1 4 ± 1 No activity No activity Compound 6 Noactivity No activity No activity 1.5 ± 0.3 Compound 7 No activity 9.5 ±2   No activity  7.6 ± 0.01

Example 7 Nuclear Receptor Selectivity Profile

Using the AlphaScreen assay, the selectivity of a compound of theinvention against the following nuclear receptors involved in themetabolic pathways can be evaluated: LXRP, PXR, CAR, PPARα, PPARα,PPARγ, RAR, RARα, VDR, TR, PR, RXR, GR, and ER.

Compounds L1, 3, 5, 6 and 7 were tested against the panel of availablenuclear receptor in both agonist and antagonist mode. Neither compoundactivated any of the receptors in agonist (dose response to 200 μM) orantagonist mode (fixed concentration at 10 μM).

Example 8 FXR Target Gene Panel

To evaluate the capacity of a compound of the invention to modulate FXRtarget genes, quantitative RT-PCR assays are performed. HepG2 cells areselected as a relevant cell line to determine whether a compound of theinvention can regulate the endogenous FXR genetic network. The abilityof a compound of the invention to induce FXR target genes is assessed byisolating total RNA from cells treated overnight with 1 μM of compoundsA, B, and a compound of the invention. Compound A is established as apotent FXR selective agonist and compound B is established as a dualpotent FXR/TGR5 agonist.

FXR regulates the expression of several target genes involved in BAhomeostasis. Briefly, FXR plays a central role in several metabolicpathways, including i.e., lipid metabolism, bile-acids metabolism, andcarbohydrate metabolism. Regarding gene expression profiling, the genesencoding proteins involved in lipid metabolism include, e.g., APOCII,APOE, APOAI, SREBP-1C, VLDL-R, PLTP, and LPL; the genes encodingproteins involved in bile-acids metabolism include, e.g., OSTα/β, BSEP,MRP2, SHP, CYP7A1, FGF19, SULT2A1, and UGT2B4; and the genes encodingproteins involved in carbohydrate metabolism include, e.g., PGCla,PEPCK, and GLUT2. FXR target genes: BSEP, SHP, OSTβ and CYP7A1 wereevaluated following stimulation of Compounds 3, 5, 6, and 7 on HepG2cells for 18 hours. Compound L1 was used as a control. Compounds 3, 5,6, and 7 significantly bind to FXR in hepatic cells modulating FXRtarget genes.

Example 9 In-Vitro Cytotoxicity

To evaluate in-vitro cytotoxicity of a compound of the invention, twodifferent assay methods are employed. The assays evaluate cell viabilityby measuring ATP levels and cytotoxicity by measuring LDH release.Adenosine Triphosphate (ATP) nucleotide represents the source of energyat the basic molecular level, as it is a multifunctional molecule thatis used in every cell as a coenzyme and is an integral part of themitochondrial DNA (Kangas, et al., Medical Biology, 1984, 62, 338-343;Crouch, et al., J Immunol. Methods, 1993, 160, 81-88; and Petty, et al.,J Biolumin. Chemilumin. 1995, 10, 29-34). It has been called the“molecular unit of currency” when it comes to intracellular energytransfer. This is to ensure the important role of ATP in metabolism anda drop in ATP content is the first step in revealing cellular damage(Storer, et al., Mutation Research, 1996, 368, 59-101; and Cree andAndreotti, Toxicology In-Vitro, 1997, 11, 553-556).

An additional method to determine the viability of cells is to detectthe integrity of the membrane that defines the cellularcompartmentalization. Measuring the leakage of components out of thecytoplasm, in damaged cell membranes, indicates loss of membraneintegrity, and LDH release is the method used to determine commontoxicity in cells. HepG2 cells are treated with a compound of theinvention, and serial dilutions are performed. LCA dilutions are addedto the plated cells as assay controls together with no-cell anduntreated cells. The assay is performed in triplicate for each testcompound concentration.

Cell viability was determined as a measure of intracellular ATP relatedto the time of exposure and concentration of the test compounds(Sussman, Promega Cell Notes, Issue 3, 2002). Data are provided inTables 5A and 5B.

TABLE 5A In vitro Cytotoxicity of Compounds 3 and 5 ATP Content EC₅₀(μM) Compound Ref Tamoxifen EC₅₀ 49 ± 9 μM Compound 3 No toxicity (100%living cells) Compound 5 No toxicity (100% living cells) Compound A* 230Compound B* 800 * Rizzo et al., Mol. Pharm. 2010, 78, 617-630.

TABLE 5B In vitro Cytotoxicity of Compounds 6 and 7 (LDH release and ATPcontent relative to Compounds 6 and 7 stimulation on HepG2. MembraneIntegrity ATP (LDH measure) Content Compound LC₅₀ (μM) EC₅₀ (μM)Tamoxifen 35 ± 10 20 ± 5 LCA 100 ± 5   75 ± 5 Compound 6 No toxicity Notoxicity Compound 7 No toxicity No toxicity Compound L1 No toxicity Notoxicity (100% living cells)

Tamoxifen was used as a positive control of the assay and LCA was usedas a reference.

Example 10 CYP4150 Screening

To evaluate the potential of a compound of the invention for drug-druginteractions, the six main CYP450 isoforms (CYP1A2, CYP2C9, CYP2C19,CYP2D6, CYP2E1, CYP3A4) are investigated (Obach, et al., J Pharmacol.Exp. Ther, 2006, 316, 336-348).

To determine interaction between a compound of the invention andcytochrome P450 enzymes, the compound of the invention is analyzed byits capacity to inhibit (or not) the production of a fluorescent signal,using recombinant CYP450 proteins (baculosomes; Invitrogen), substratesand inhibitors (Bidstrup, et al., Br J Clin. Pharmacol, 2003, 56,305-14). As a positive control, a selective inhibitor for each CYP450isoform is tested in the same plate.

TABLE 6 CYP450s Inhibition (test against the 6 major isozymes) CompoundCompound Compound Compound Compound 3 5 6 7 L1 CYP450 IC₅₀ (μM) IC₅₀(μM) IC₅₀ (μM) IC₅₀ (μM) IC₅₀ (μM) CYP1A2 >10 >10 >10 >10 >10CYP3A4 >10 >10 >10 >10 >10 (Green substrate) CYP3A4 >10 >10 >10 >10 >10(Blue substrate) CYP2C9 >10 >10 >10 >10 >10 CYP2C19 >10 >10 >10 >10 >10CYP2D6 >10 >10 >10 >10 >10 CYP2E1 >10 >10 >10 >10 >10

Example 11 Human ERG Potassium Channel

To determine on channel function, the Predictor™ hERG FluorescencePolarization assay is employed as it provides an efficient method for aninitial determination of the propensity of test compounds to block thehERG channel (Dorn, et al, J Biomol. Screen, 2005, 10, 339-347). Theassay is based on the assumption that the hERG potassium channelactivity contributes to the resting membrane potential in permanentlytransfected cells, and thus a block of hERG channels should result in adepolarization of the cell membrane. The assay is designed to identifypotential hERG channel blockers by producing data that accuratelycorrelates with patch-clamp electrophysiology studies. Results from thePredictor™ assay demonstrate a high correlation with those obtained frompatch clamp techniques (Dorn, et al., J Biomol Screen, 2005, 10,339-347).

Membrane preparations from Chinese hamster ovary cells stablytransfected with hERG potassium channel are used to evaluate thepotential inhibitory effect of a compound of the invention on thischannel using the Predictor™ fluorescence polarization assay. Reductionof membrane polarization as a result of inhibition of the hERG potassiumchannel is directly correlated with a reduction of the fluorescencepolarization (FP).

The assay is performed in triplicate by using a 16-point dose-responseof test compound and the positive controls E-4031 and Tamoxifen. An IC₅₀of 15 nM (AmP=163) for E-4031 and 1,4 μM (ΔητP=183) for Tamoxifen areobtained. An assay window more than 100 mP (millipolarization) isconsidered good. The non-linear regression curves are obtained byGraphPad Prism (GraphPad Software Inc.) analysis, to calculate the IC₅₀values.

TABLE 7 Human ERG potassium channel inhibition hERG inhibition CompoundIC₅₀ (μM) Compound 3 >100 Compound 5 >100 Compound 6 >100 Compound7 >100 Compound L1 >100 Compounds L1, 3, 5, 6 and 7 did not inhibit hERGpotassium channel.

Example 12 Physiochemical Properties

Physiochemical properties of a compound of the invention such as watersolubility, critical micellar concentration, surface tension, andLogP_(A) were determined using methods known in the art. Data areprovided in Table 8.

TABLE 8 Physiochemical Properties Bile Acid CMC^((a)) Derivative (mM)LogP_(A−) ^((b)) Compound 3 12.5 0.12 Compound 5 8.5 0.61 Compound 6 281.7 Compound 7 — 2.0 Compound L1 15.8 0.84 Compound A 2.9 2.5 Compound B1.3 2.0 Compound C 2 1.4 Compound D — 2.9 Compound E 5.9 1.6 ^((a))CMC:Critical Micellar Concentration determined in 0.15 M NaCl water solution^((b))LogP_(A−): 1-octanol-water species partition coefficient of thestudied bile acids as ionized species

1. A compound of formula I:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof, wherein: R¹ is OH, alkoxy, halogen, or oxo; R² and R³ are eachindependently H, OH, halogen, or alkyl optionally substituted with oneor more halogen or OH, or R² and R³ taken together with the carbon atomto which they are attached form a carbonyl; R⁴ is H, halogen, alkyloptionally substituted with one or more halogen or OH, alkenyl, oralkynyl; R⁵ and R⁶ are each independently H, OH, OSO₃H, OCOCH₃, OPO₃H₂,or halogen, or R⁵ and R⁶ taken together with the carbon atom to whichthey are attached form a carbonyl; R⁷ is OH, OSO₃H, SO₃H, OSO₂NH₂,SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, C(O)NHOH, tetrazolyl, oxadiazolyl,thiadiazolyl, 5 -oxo-1,2,4-oxadiazolyl, 5-oxo-1,2,4-thiadiazolyl,oxazolidine-dionyl, thiazolidine-dionyl, 3-hydroxyisoxazolyl,3-hydroxyisothiazolyl, or 2,4-difluoro-3-hydroxyphenyl; R⁸, R⁹, and R¹⁰are each independently H, OH, halogen, or alkyl optionally substitutedwith one or more halogen or OH, or R⁸ and R⁹ taken together with thecarbon atoms to which they are attached form a 3- to 6-memberedcarbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selectedfrom N, O, and S, or R⁹ and R¹⁰ taken together with the carbon atoms towhich they are attached form a 3- to 6-membered carbocyclic orheterocyclic ring comprising 1 or 2 heteroatoms selected from N, O, andS; m is 0, 1, or 2; n is 0 or 1; p is 0 or 1; and

is a single or double bond, provided that when each

is a single bond, the sum of m, n, and p is 2, R¹ is OH, and R⁸, R⁹, andR¹⁰ are each H, then R⁷ is not CO₂H.
 2. The compound of claim 1, whereinthe compound is of formula Ia:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.
 3. The compound of claim 1, wherein the compound is of formulaIb or Ic:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.
 4. The compound of claim 1, wherein R¹ is OH, alkoxy, or oxo.5-8. (canceled)
 9. The compound of claim 1, wherein R³ is H. 10-11.(canceled)
 12. The compound of claim 1, wherein R⁶ is OH or H. 13.(canceled)
 14. The compound of claim 1, wherein the compound is offormula Id:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.
 15. The compound of claim 1, wherein the compound is of formulaIe:

or a pharmaceutically acceptable salt, solvate, or amino acid conjugatethereof.
 16. The compound of claim 1, wherein R⁷ is OH, OSO₃H, SO₃H,OSO₂NH₂, SO₂NH₂, OPO₃H₂, PO₃H₂, CO₂H, or C(O)NHOH. 17-23. (canceled) 24.The compound of claim 1, wherein R⁷ is tetrazolyl, oxadiazolyl,thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl, 5-oxo-1,2,4-thiadiazolyl,oxazolidine-dionyl, thiazolidine-dionyl, 3-hydroxyisoxazolyl,3-hydroxyisothiazolyl, or 2,4-difluoro-3-hydroxyphenyl.
 25. (canceled)26. The compound of claim 1, wherein R² is OH. 27-28. (canceled)
 29. Thecompound of claim 1, wherein R⁵ is OH or H. 30-34. (canceled)
 35. Thecompound of claim 1, wherein R⁴ is methyl, ethyl, or propyl. 36-40.(canceled)
 41. The compound of claim 1, wherein R⁴ is in the α-position.42. The compound of claim 1, wherein R¹ is in the β-position.
 43. Thecompound of claim 1, wherein the compound is selected from:


44. A salt of the compound of claim
 1. 45. The compound of claim 44,wherein R⁷ is OSO₃ ⁻.
 46. The compound of claim 45, wherein R⁷ is OSO₃⁻Na⁺. 47-51. (canceled)
 52. A pharmaceutical composition comprising thecompound of claim 1 and a pharmaceutically acceptable carrier orexcipient.
 53. A method of treating or preventing a disease or conditionin a subject in need thereof comprising administering an effectiveamount of the compound of claim 1 or a pharmaceutically acceptable salt,solvate, or amino acid conjugate thereof, and wherein the disease orcondition is mediated by FXR.
 54. The method of claim 53, wherein thedisease is selected from cardiovascular disease, chronic liver disease,lipid disorder, gastrointestinal disease, renal disease, metabolicdisease, cancer, and neurological disease. 55-61. (canceled)
 62. Acompound of claims 1 wherein R₇ is OH.
 63. The compound of claim 62wherein the compound is selected from:


64. (canceled)